EPICENTRE Enzyme Catalog-Beta/Int'l version

Enzyme Catalog 

Your trusted source for specialty molecular biology enzymes

Introduction 

EPICENTRE Profile 

Our expertise in producing enzymes 

EPICENTRE Biotechnologies develops, 

manufactures, and sells high-quality enzyme 

systems for life science research. Located in 

Madison, Wisconsin, EPICENTRE was founded 

in 1987, and now occupies a state-of-the art 

72,000-s.f. building. EPICENTRE is well-known 

for its unique expertise in making a broad 

range of enzymes for molecular biology, many 

of which are unique to EPICENTRE. EPICENTRE 

products are available directly in the United 

States and internationally through authorized 

distributors 

Highest enzyme purity in the market 

EPICENTRE takes pride in its optimized and 

proprietary protein production, purification and 

quality control procedures that contribute to the 

extremely high purity of our enzymes products. 

Unit definition 

A significant number of commercially available 

enzymes have different unit definitions from 

different suppliers. Unit definition is directly 

related to the protocol on how to use an enzyme 

and to its actual cost. Some of our enzymes’ unit 

definition makes one EPICENTRE unt equivalent 

to multiple units of the same enzymes from 

some other suppliers. Please visit www.EpiBio. 

com or call our Tech Support at 1-800-284- 

8474 if you have questions on our enzyme unit 

definition. 

Tag free 

EPICENTRE’s optimal and proprietary enzyme 

production procedure ensures that no tags 

are used in the production of our final enzyme 

products. If you are concerned about various 

tags affecting your research applications, 

EPICENTRE is your choice of enzyme source. 

Animal Product Free 

We also produce a range of animal productfreeenzymes. 

Please visit www.EpiBio.com or 

call Customer Service at 1-800-274-8474 for 

more information 

Bulk and custom offerings 

EPICENTRE always tries to accommodate 

our customers’ needs for your specific 

applications. If you need enzymes with bulk 

quantitities, and/or custom formulations (e.g. 

higher concentrations), please inquire with our 

Customer Service at 1-800-274-8474. 

Limited use license 

EPICENTRE’s products are labeled for research 

or laboratory use only. For any orther use please 

inquire through our toll free number 1-800-284- 

8474. 

NOTICE TO PURCHASER: LIMITED LICENSE 

*Use of MasterAmp AmpliTherm DNA Polymerase, MasterAmp Taq DNA Polymerase, MasterAmp Tfl DNA Polymerase, or MasterAmp Tth DNA 

Polymerase is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,079,352, 5,789,224, 5,618,711, 

6,127,155 and claims outside the US corresponding to US Patent No. 4,889,818. The purchase of this product includes a limited, non-transferable immunity 

from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under any other patent 

claim (such as the patented 5´ Nuclease Process claims in US Patents Nos. 5,210,015 and 5,487,972), no right to perform any patented method, and no 

right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activities for a fee or other commercial 

consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a 

separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 

Lincoln Centre Drive, Foster City, California 94404, USA. 

Use of MasterAmp PCR Enhancer DNA Polymerase Reactions, including, but not limited to use for PCR or DNA Sequencing, is covered by U.S. 

Patent No. 6,270,962, European Patent No. 0742838, German Patent No. DE4411588C1, and other issued or pending applications in the U.S. and other 

countries that are either assigned or exclusively licensed to EPICENTRE. These products are accompanied by a limited non-exclusive license for the purchaser 

to use the purchased products solely for life science research. Contact EPICENTRE for information on licenses for uses in diagnostics or other fields. 

2 

techhelp@EpiBio.com • www.EpiBio.com

T7, T3, and SP6 RNA Polymerases 

Produce defined RNA by in vitro transcription of 

double-stranded DNA that is downstream of the 

respective RNA polymerase promoter. 

• Extremely high promoter specificity. 

Applications 

• RNA synthesized can be used as a 

hybridization probe, anti-sense RNA, a 

ribozyme, a template for in vitro translation, 

as a precursor mRNA for splicing or other 

processing studies, or to make dsRNA for 

RNA interference or gene silencing. 

• Synthesis of RNA for nucleic acid 

amplification methods or gene expression 

studies. 

Catalog No. Conc. Size Price $ 

T7 RNA Polymerase 

T7905K 50 U/µl 5,000 U 

T7925K 50 U/µl 25,000 U 

TM910K 200 U/µl 10,000 U 

TM925K 200 U/µl 25,000 U 

TH950K 1,000 U/µl 50,000 U 

TU950K 2,500 U/µl 50,000 U 

(Enzyme only. Transcription Buffer is not included.) 

T3 RNA Polymerase 

T4905K 50 U/µl 5,000 U 

T9050K 50 U/µl 50,000 U 

TM4910K 200 U/µl 10,000 U 

TH4950K 1,000 U/µl 50,000 U 


SP6 RNA Polymerase 

S4905K 50 U/µl 5,000 U 

S4950K 50 U/µl 50,000 U 

SM910K 200 U/µl 10,000 U 


Transcription Buffer Package 

BP1001 - 1 pkg 

Includes 5 ml of 5X Transcription Buffer and 2.5 ml of 100 

mM DTT. 

*Greatest range of enzyme concentrations available from 50 

U/µl to 2,500 U/µl. 

**For kits for in vitro transcription, please visit 

www.EpiBio.com/ivt.asp. 

RNA Polymerases and Replicases 

T7 R&DNA Polymerase and SP6 R&DNA Polymerase 

Recombinant enzymes having single-base 

active-site mutations in the respective T7 or 

SP6 RNA Polymerase gene. These active-site 

mutations enable the corresponding T7 or 

SP6 R&DNA Polymerase to incorporate 2’- 

deoxyribonucleoside triphosphates (dNTPs) into 

full-length transcripts much more efficiently 

than the corresponding wild-type enzymes, 

while retaining the same catalytic activity for 

incorporation of canonical NTPs and the same 

high promoter specificity. 

Applications 

• Synthesis of “RNA” transcripts of mixed 

rNMP/2’-dNMP or rNMP/2’-modified-NMP 

composition. 

• Synthesis of modified “RNA” transcripts that 

are resistant to RNase A-type RNases. 

T7 & SP6 R&DNA Polymerases, & DuraScribe T7 & SP6 

Transcription Kits to synthesize nucleic acids with non-canonical 

bases or for partial ribosubstitution are covered by U.S. Patents 

5,849,546; 6,107,037; or 6,596,494, and other patents issued 

or pending. These products are accompanied by a limited nonexclusive 

license for the purchaser to use the purchased product(s) 

solely for life science research except the development of 

therapeutics. Contact EPICENTRE concerning licenses for other uses. 


T7 R&DNA Polymerase 

D7P9201K 50 U/µl 1,000 U 

D7P9205K 50 U/µl 5,000 U 

Contents: T7 R&DNA Polymerase, 5X Reaction Buffer, and 

100 mM DTT. 

SP6 R&DNA Polymerase 

D6P9301K 50 U/µl 1,000 U 

D6P9305K 50 U/µl 5,000 U 

Contents: SP6 R&DNA Polymerase, 5X Reaction Buffer, 

and 100 mM DTT. 

*For kits for in vitro transcription, please visit 


E. coli RNA Polymerase Core Enzyme and Sigma-Saturated Holoenzyme 

Both enzyme preparations are isolated from the 

rifampicin-sensitive strain BL21. EPICENTRE 

is the only company that offers purified Core 

Enzyme, which has no detectable sigma subunit, 

and 100% Sigma-Saturated (σ 70 )-Holoenzyme. 

Applications 

• The Core Enzyme is useful in studying 

mechanisms of transcription initiation, 

since it will not initiate specific transcription 

at promoter sequences on bacterial or 

bacteriophage DNA due to a lack of sigma 

factor. 

• The sigma-saturated Holoenzyme is very 

efficient in specifically transcribing a variety 

Derived from the thermophilic bacterium, 

Thermus thermophilus, this RNA Polymerase 

is the only commercially-available RNA 

of double-stranded DNA templates containing 

promoters. 

FIG 1. Subunit patterns of 

E. coli RNA Polymerase 

Core Enzyme and 

- β´, β 

- σ Holoenzyme preparations. 

Equivalent amounts of each 

- α enzyme were separated by 

electrophoresis on an SDS 

15% polyacrylamide gel, 

stained with Coomassie Blue, 

and dried. The sigma-70 

- ω subunit is 100% saturating in 

the Holoenzyme, but is absent 

in the Core Enzyme. 

Holo Core 

polymerase that is stable and has optimal 

activity at temperatures above 65°C. 

Catalog No. Conc. Size 

E. coli RNA Polymerase Core Enzyme 

C90100 - 100 U 

C90250 - 250 U 

E. coli RNA Polymerase Holoenzyme 

(Sigma-Saturated) 

S90050 - 50 U 

S90100 - 100 U 

*For kits for in vitro transcription, please visit 


Thermus Thermostable RNA Polymerase 


Thermus Thermostable RNA Polymerase 

T90050 - 50 U 

T90100 - 100 U 

www.EpiBio.com • techhelp@EpiBio.com 

3

ScriptCap m7G Capping System 

RNA Capping and Tailing Enzymes 


ScriptCap m 7 G Capping System 

SCCE0610 - 10 Reactions 

SCCE0625 - 25 Reactions 

Contents: ScriptCap Capping Enzyme, 10X Capping Buffer, 

ScriptGuard RNase Inhibitor, 20 mM SAM, 10 mM GTP, and 

RNase-Free Water. 

*For complete kits designed to produce capped and tailed 

RNA in vitro, please visit www.EpiBio.com/capping.asp. 

ScriptCap 2’-O-Methyltransferase 

Based upon the tri-functional Vaccinia Virus 

capping enzyme (VCE), this system is designed 

to build the Cap 0 structure found on the 5´ 

end of most eukaryotic mRNA molecules. This 

enzyme system is identical to the Vaccinia 

guanylyltransferase sold by other vendors. 

Applications 

• In vitro production of capped RNA for in vivo 

or in vitro translation.* 

• Analysis of 5´ ends of RNA transcripts. 

Figure: (A) Denaturing Polyacrylamide gel and (B) 

autoradiograph of a ScriptCap Capping system 

reaction. Lane 2 shows the uncapped transcript 

without Vaccinia capping enzyme (VCE). Lane 

3 includes the VCE and 14C SAM. VCE has clearly 

transferred the guanine base (Lane 3A) and the 14C 

containing methyl group (Lane 3B). 


ScriptCap 2’-O-Methyltransferase 

SCMT0610 - 10 Reactions 

SCMT0625 - 25 Reactions 

Contents: ScriptCap 2’-O-Methyltransferase, 10X Capping 

Buffer, and 20 mM SAM. 

* For complete kits designed to produce capped and tailed 

RNA in vitro, please visit www.EpiBio.com/capping.asp. 

The ScriptCap 2’-O-Methyltransferase 

Enzyme is derived from the Vaccinia virus and 

methylates the penultimate nucleotide in a 

capped eukaryotic mRNA transcript, converting 

a Cap 0 transcript into the natural Cap 1 mRNA 

structure. This methylation results in an up to 

50% increase in the in vivo translation efficiency 

when compared to Cap 0 mRNA. 

The enzyme is active on both the natural 

Cap 0 structure and several different Cap 0 

dinucleotide analogs used in co-transcriptional 

capping kits. Each reaction is capable of 

methylating 60 ug of Cap 0 RNA in 30 minutes. 

Applications 

Table 1. Various different forms of capped and tailed mRNA were transfected into HeLa cells and assayed 

for luciferase activity. Data is normalized to the Cap 0, poly(A) transfection results. The complete Cap 1 mRNA 

exhibited up to 50% higher in vivo translation efficiency when compared to the various Cap 0-type structures 

currently used in most transfections. 

Methylation of Cap 0 RNA to the Cap 1 form 

Means of Cap 0 Production 

ScriptCap 

2’-O-Methyltransferase 

Treatment 

Final mRNA Cap 

Structures Formed 

Translation Efficiency 

relative to non-treated 

mRNA 

No RNA no none 0% 

ScriptCap m7G Capping System no 

m7GpppN 

100% 

(Capping Enzyme) 

(Cap 0) 

ScriptCap m7G Capping System 

(Capping Enzyme) 

AmpliCap-Max High Yield 

Message Maker Kit 

(Standard Cap Analog) 

AmpliCap-Max High Yield 

Message Maker Kit 

(Standard Cap Analog) 

AmpliScribe T7-Flash 

Transcription Kit 

(ARCA Cap Analog) 

AmpliScribe T7-Flash 

Transcription Kit 

(ARCA Cap Analog) 

yes 

m7Gppp[m2’-O]N 

(Cap 1) 

no m7GpppN * 

(Cap 0) 

yes m7Gppp[m2’-O]N * 

(Cap 1) 

no m27, 3´-OGpppN * 

(Cap 0) 

yes m27, 3´-OGppp[m2’-O]N * 

(Cap 1) 

147% 

100% 

148% 

100% 

128% 

A-Plus Poly(A) Polymerase 

4 


A-Plus Poly(A) Polymerase Tailing Kit 

50 

PAP5104H 4 U/µl Reactions 

(400 U) 

Contents: A-Plus Poly(A) Polymerase, A-Plus 10X 

Reaction Buffer, 10 mM ATP, and Sterile RNase-Free Water. 

A-Plus Poly(A) Polymerase uses ATP as a 

substrate for template-independent addition of 

adenosine monophosphate to the 3´-hydroxyl 

termini of RNA molecules. The A-Plus Poly(A) 

Polymerase Tailing Kit provides the enzyme 

and other reagents to quickly and easily add a 

“poly(A) tail” to the 3´-end of any RNA. 

Applications 

• Addition of a poly(A) tail to RNA synthesized 

in vitro to increase RNA stability and enhance 

its in vivo translation after transfection or 

microinjection into eukaryotic cells. 

• Addition of a poly(A) tail to RNA molecules to 

provide a priming site for synthesis of firststrand 

cDNA using a primer with poly(dT) on 

its 3´-end. 

• Cloning of DNA encoding RNA molecules of 

unknown or multiple sequences by adding 

a poly(A) tail that can anneal to a T-tailed 

vector. 

• Synthesis of polyadenylated RNA for 

nucleic acid amplification methods or gene 

expression studies. 

• 3´-End-labeling of RNA with radioactive A 

residues. 

• Quantifying mRNA. 


Product Name 

Properties of Mesophilic DNA Polymerases 

Activity 

5´→3´ exonuclease 3´→5´ exonuclease Nick translation Heat Inactivation a Strand displacement 

DNA Polymerase I, E. coli + ++ + 75°C 20 minutes - 

Klenow DNA Polymerase - ++ - 75°C 20 minutes + 

Exo-Minus Klenow DNA 

Polymerase 

RepliPHI Phi29 DNA 

Polymerase 

- - - 75°C 20 minutes + 

- ++ - 65°C 10 minutes ++++ 

T4 DNA Polymerase - +++ - 75°C 20 minutes - 

T7 DNA Polymerase, 

Unmodified 

a 

Indicated treatment results in complete inactivation under standard reaction conditions 

- +++ - 75°C 20 minutes - 

DNA Polymerases 

This DNA-dependent DNA polymerase contains 

both 5’→3’and 3´→5´ exonuclease activities. 

The 5´→3´ exonuclease activity enables the 

enzyme to use nicks and gaps in the DNA as 

starting points for labeling the DNA by nick 

translation. 

Applications 

• Generate labeled DNA probes by nick 

translation. 

• Second strand cDNA synthesis 

• In vitro synthesis of DNA. 

DNA Polymerase I, E. coli 


DNA Polymerase I, E. coli 

DP02250 10 U/µl 250 U 

DP021K 10 U/µl 1000 U 

Derived from E. coli DNA polymerase I, this 

large fragment, DNA-dependent enzyme has 

5´→3´ polymerization and 3´→5´ exonuclease 

activities, but lacks 5´→3´ exonuclease 

activity. Klenow DNA polymerase blunt ends 

doublestranded DNA with singlestranded 

overhangs. The 3´→5´ exonuclease activity 

removes 3´ overhangs and the 5´→3´ 

polymerization activity fills in 5´ overhangs. 

Applications 

• Random primer labeling of DNA. 

• DNA sequencing. 

• Second-strand cDNA synthesis. 

• Strand displacement amplification. 

Klenow DNA Polymerase 


Klenow DNA Polymerase 

KP04061K - 1000 U 

This DNA-dependent DNA polymerase lacks 

both the 5´→ 3´ and 3´→5´ exonuclease 

activities of E. coli DNA Polymerase I from which 

it is derived. 

Applications 

• Random primer labeling of DNA. 

• DNA sequencing. 

• Second-strand cDNA synthesis. 

• Strand displacement amplification. 

Exo-Minus Klenow DNA Polymerase 


Exo-Minus Klenow DNA Polymerase 

KL04011K 20 U/µl 1000 U 

KL06041K 50 U/µl 1000 U 

RepliPHI Phi29 DNA Polymerase (φ29 DNA 

Polymerase) is a highly processive enzyme with 

exceptional strand displacement activity. The 


enzyme also contains a 3´→5´ exonuclease 

activity that enables proofreading capability. Its 

specific activity is 1 x 10 6 U/mg. 

RepliPHI Phi29 DNA Polymerase 


RepliPHI Phi29 DNA Polymerase (enzyme only) 

PP031010 

1 µg/µl 10 µg 

(1,000 U/µl) (10,000 U) 

PP040110 

0.1 µg/µl 10 µg 

(100 U/µl) (10,000 U) 

RepliPHI Phi29 Reagent Set 

(includes enzyme, buffer, dNTPs, DTT) 

RH031110 

Enzyme: 

1 µg/µl 

(1,000 U/µl) 


10 µg 

(10,000 U) 

RH040210 


0.1 µg/µl 

(100 U/µl) 


10 µg 

(10,000 U) 

RepliPHI Phi29 Polymerase Dilution Buffer 

RPB04041 - 1 ml 

5

T4 DNA Polymerase 



T4 DNA Polymerase 

D0602H - 200 U 

D0605H - 500 U 

Contains both a template-directed 5´→3´ 

DNA polymerase activity and a potent 3´→ 5´ 

exonuclease activity. 

Applications 

• Conversion of 5´- and 3´-protruding DNA 

termini to blunt ends. 

• Cloning of PCR fragments: Treatment of PCR 

products containing 3´-A overhangs with T4 

DNA Polymerase and dNTPs produces blunt 

ends. 

• Production of site-specific mutations: 

this enzyme can be used for site-specific 

mutagenesis by primer extension of 

“mutated” oligonucleotides hybridized to 

single-stranded DNA templates. 

• Labeling of 3´-termini of DNA molecules and 

synthesis of strand-specific probes using the 

exonuclease and polymerase activities of T4 

DNA Polymerase. 

T7 DNA Polymerase, Unmodified 


T7 DNA Polymerase, Unmodified 

D07250 10 U/µl 250 U 

D07500 10 U/µl 500 U 

Contains both a template-directed 5´→ 3´ 

DNA polymerase activity and a potent 3´→ 

5´ exonuclease activity. The unmodified form 

of T7 DNA Polymerase is different from T7 

DNA polymerase preparations from which 

exonuclease activity has been removed. The 

enzyme is a tightly-bound complex of T7 

phage-encoded gene 5 protein and E. coli hostencoded 

thioredoxin. 

Highly processive and synthesizes long 

stretches of DNA before dissociating from the 

template. The rate of elongation is also much 

faster than that of most other DNA polymerases. 

Applications 

• Primer extension of long DNA molecules. 

• Conversion of 5´- and 3´-protruding ends to 

blunt ends. 

• Labeling of 3´-ends of DNA. 

• In situ detection of DNA fragmentation 

associated with apoptosis. 

Product Name 

Reverse 

transcriptase 

Properties of Thermophilic DNA Polymerases 

5´→3´ 

exonuclease 

3´→5´ 

exonuclease 

rBst DNA Polymerase a + + -

Bst DNA Polymerase, Large Fragment (IsoTherm DNA Polymerase) 

Derived from the DNA pol I gene of 

the thermophilic bacterium Bacillus 

stearothermophilus (Bst) altered to remove the 

5´→ 3´ DNA exonuclease activity. Having optimal 

DNA polymerase activity at 65°C, the enzyme 

is suitable for high-temperature isothermal 

sequencing of DNA. Like the Klenow fragment 

of E. coli DNA Polymerase I, this enzyme has 

strong strand displacement activity. It also has 

thermostable RNA-dependent DNA polymerase 

(i.e., reverse transcriptase) activity. 

Ability to synthesize through regions of high GC 

content or difficult secondary structure at high 

temperatures. 

Its high rate of DNA synthesis permits use 

of nanogram quantities of template under 

isothermal conditions. 

Applications 

• Amplification assays and other assays 

involving continuous displacement of the DNA 

strand concomitant with high-temperature 

DNA synthesis. 

• First-strand cDNA synthesis from primed RNA 

templates. 

• High-temperature isothermal DNA 

sequencing. 


rBst DNA Polymerase, Large Fragment 

(IsoTherm DNA Polymerase) 

BL901K 5 U/µl 1,000 U 

BL1805K 50 U/µl 5,000 U 

BL1950K 50 U/µl 50,000 U 

Enzyme only. 


MasterAmp AmpliTherm DNA Polymerase* 

This proprietary recombinant thermostable 

DNA polymerase for use in PCR has optimal 

DNA synthetic activity at temperatures above 

70°C and can be used at temperatures up to 

95°C. It lacks both 5´→ 3´ structure-dependent 

exonuclease activity like that found in Taq 

DNA polymerase and 3´→ 5´ proofreading 

exonuclease activity found in some other DNA 

polymerases. 

Provided with the MasterAmp PCR Enhancer, 

which increases the probability of obtaining the 

desired amplification product, the reproducibility 

of PCR, and improves the consistency of PCR 

product yields in multiplex PCR. 

Applications 

• PCR amplification of DNA templates. 

• Multiplex PCR. 


MasterAmp AmpliTherm DNA Polymerase 

AT72250 5 U/µl 250 U 

Includes 10X PCR Buffer, 25 mM MgCl 2 

, and MasterAmp 

10X PCR Enhancer. 

MasterAmp AmpliTherm DNA Polymerase 

(Enzyme Only) 

AT72250N 5 U/µl 250 U 

For use with MasterAmp PCR PreMixes. 

MasterAmp Taq DNA Polymerase* 

This thermostable DNA polymerase derived 

from Thermus aquaticus has optimal activity 

at temperatures above 70°C. It has an intrinsic 

5´→ 3´ structure-dependent exonuclease 

activity, but lacks 3´→ 5´ proofreading 

exonuclease activity. Reliable activity, specificity, 

and reproducibility in PCR in conjunction with 

the MasterAmp PCR Enhancer Technology. 

Applications 

• PCR and Multiplex PCR amplification of DNA 

templates. 


MasterAmp Taq DNA Polymerase 

Q82250 5 U/µl 250 U 

Q8201K 5 U/µl 1,000 U 

Includes 10X PCR Buffer, 25 mM MgCl 2 

, and MasterAmp 


MasterAmp Taq DNA Polymerase (Enzyme Only) 

Q82250N 5 U/µl 250 U 


MasterAmp Taq PCR Core Kit 

MCQ74200 - 200 Reactions 

Contents: MasterAmp Taq DNA Polymerase, 10X PCR 

Buffer, MasterAmp 10X PCR Enhancer, dNTP Mix, 2.5 

mM each, 25 mM MgCl 2 

, Enzyme Dilution Buffer, Control 

Template and Primers Mix. 

MasterAmp Tfl DNA Polymerase* 

Derived from the thermophilic bacterium Thermus 

flavus, this is a recombinant DNA polymerase 

with good thermostability (to ~95°C) and 

processivity (with 15 kb PCR products reported). 

MasterAmp PCR Enhancer Technology 

substantially increases the probability of 

obtaining the desired amplification product and 

the reaction-to-reaction consistency, and greatly 

improves the consistency of PCR product yields in 

multiplex PCR. 

Applications 

• PCR and Multiplex PCR amplification of DNA 

templates. 

• Produces PCR products up to 15 kb. 

• Adaptable to high-throughput PCR formats. 


MasterAmp Tfl DNA Polymerase 

F72250 1 U/µl 250 U 

F7201K 1 U/µl 1,000 U 

Includes 20X PCR Buffer, 25 mM MgCl2, and MasterAmp 


MasterAmp Tfl DNA Polymerase (Enzyme Only) 

F72250N 1 U/µl 250 U 


MasterAmp Tth DNA Polymerase* 


This recombinant DNA enzyme from Thermus 

thermophilus has both DNA-dependent and 

RNA-dependent (i.e., reverse transcriptase) DNA 

polymerase activities up to ~95°C. High reaction 

temperatures can reduce nonspecific priming 

and template secondary structure. Provided with 

MasterAmp PCR Enhancer Technology. 

Has both reverse transcriptase and DNAdependent 

DNA polymerase activity under the 

same reaction conditions. 

Improved PCR of RNA and DNA templates having 

a high degree of secondary structure. 

Applications 

• PCR amplification of DNA 

• 1-step RT-PCR of RNA templates. 


MasterAmp Tth DNA Polymerase 

TTH72100 5 U/µl 100 U 

TTH72250 5 U/µl 250 U 

TTH7201K 5 U/µl 1,000 U 

Includes a 20X PCR Buffer (without Mg2+ or Mn2+) plus 

separate 25 mM solutions of MgCl2 and MnSO4, and 

MasterAmp 10X PCR Enhancer. 

MasterAmp Tth DNA Polymerase (Enzyme Only) 

TTH7225N 5 U/µl 250 U 


*Refer to page 2 for patent and license information. 

7

MMLV Reverse Transcriptase 

Reverse Transcriptase 


MMLV Reverse Transcriptase 1st-Strand 

cDNA Synthesis Kit 

MM070110 - 10 Reactions 

MM070150 - 50 Reactions 

Contents: MMLV Reverse Transcriptase, 10X Reaction 

Buffer, ScriptGuard RNase Inhibitor, dNTP PreMix, DTT, 

Oligo(dT)21Primer, Random 9-mer Primers, RNase-free 

Water. 

*See page XX for RNase H and related products 

The MMLV Reverse Transcriptase 1st-Strand 

cDNA Synthesis Kit is optimized for generating 

full-length first-strand cDNA from total cellular 

RNA preparations or purified polyadenylated RNA. 

• Synthesize full-length cDNA from RNA 

templates longer than 15 kb. 

• Both oligo(dT) and random primers included 

in the kit. 

• The kit includes both an oligo(dT)-containing 

and a random nonamer (9-mer) primer. 

• First-strand cDNA can be made from 

picogram amounts of total RNA. 

• A potent RNase Inhibitor is included. 

Applications 

• First-strand cDNA synthesis. 

• Production of cDNA for subsequent PCR or 

real-time PCR. 

• RT-PCR validation of gene expression data 

obtained from microarray experiments. 

• RT-PCR validation and quantification of gene 

silencing by RNA interference. 

~ 15.2 kb HERC1 mRNA 

5' AAAA(A) n 

3' 

cDNA Synthesis 

3' TTTT(T) 17 

5' 

1.3 kb PCR product 

FIG 1. The MMLV Reverse Transcriptase 1st-Strand 

cDNA Synthesis Kit produces full-length cDNA 

from mRNA longer than 15 kb. Total RNA isolated 

from HeLa cells was reverse transcribed and the cDNA 

was amplified as described in the text. A. Detection of 

the 1.3 kb PCR amplicon from near the 5´ end of the 

mRNA demonstrates full-length reverse transcription 

of HERC1 mRNA. B. Agarose gel analysis of the PCR 

reaction shows the 1.3 kb amplicon from the 5´-end 

of the mRNA. Lane M, 100 bp DNA ladder; Lane 1, no 

reverse transcriptase control reaction; Lane 2, PCR 

product from cDNA synthesized by EPICENTRE’s MMLV 

Reverse Transcriptase 1st-Strand cDNA Synthesis Kit. 

1A 

i2130703 

1B 

MonsterScript 1st-Strand cDNA Synthesis Kit 

8 


MonsterScript Reverse Transcriptase 

MSTA5110 - 10 Reactions 

MSTA5124 

24 Reactions 

The MonsterScript 1st-Strand cDNA 

Synthesis Kit is optimized for generating 

full-length first-strand cDNA from multiple 

mRNA species in samples containing total 

cellular or purified polyadenylated RNA. This kit 

uses MonsterScript Reverse Transcriptase, 

a reverse transcriptase that lacks RNase H 

activity. The enzyme’s lack of RNase H activity 

contributes to its ability to make longer cDNAs 

and more complete libraries of first-strand cDNA 

molecules. 

• Lacks RNase H, enabling improved synthesis 

of full-length cDNA even for long mRNA. 

• MonsterScript is thermostable, permitting 

reverse transcription at temperatures >50°C, 

which reduces RNA secondary structure and 

improves priming specificity. 

• MonsterScript RT PreMix contains 

optimized concentrations of dNTPS, Mg +2 

and Betaine for superior performance and 

minimal pipetting steps. 

• Betaine in the cDNA Synthesis PreMix 

reduces pausing and stops during reverse 

transcription. 

• The kit includes both an oligo(dT)-containing 

and a random nonamer primer. 

• First-strand cDNA can be made from 

picogram amounts of total RNA. 

• A potent RNase Inhibitor is included. 

Applications 

• First-strand cDNA synthesis. 

• Production of cDNA for subsequent PCR or 

real-time PCR. 

• RT-PCR validation of gene expression data 

obtained from microarray experiments. 

• RT-PCR validation and quantification of gene 

silencing by RNA interference. 

1A 

~ 15.2 kb HGNEFp532 mRNA 

5' AAAAA . . . -3' 

1.3 kb 

PCR 

—1.3 kb 

FIG 1. MonsterScript Reverse Transcriptase 

produces full-length cDNA from mRNA greater 

than 15 kb. The ≈15.2 kb HGNEFp532 mRNA 

was reverse transcribed from total HeLa RNA in a 

standard MonsterScript reaction. Two microliters of 

the reaction was used to PCR amplify a 1.3 kb region 

within 68 bases of the 5´-end of the HGNEFp532 

mRNA (1A). Agarose gel of the 1.3 kb amplicon from 

the 5´-end of the mRNA demonstrates full-length 

cDNA synthesis (1B). 

1B 

techhelp@EpiBio.com • www.EpiBio.com 

i470407ms

DNA and RNA Exonuclease 

Properties of Exonucleases and Endonucleases: Active on both DNA and RNA 

Enzyme Substrate Activity Products Applications 

Terminator 

5´-Phosphate- 

Dependent 

Exonuclease 

ssDNA 

or 

ssRNA 

DNA and RNA Endonucleases 

Mung Bean 

Nuclease 

OmniCleave 

Endonuclease 

ssDNA 

or 

ssRNA 

single- and 

double-stranded 

DNA and RNA 

5´→ 3´ exonuclease that 

digests ssDNA or ssRNA with 

5´-phosphorylated ends, but 

not with 5´-hydroxylated ends. 

Single-strand-specific 

endonuclease. 

Endonuclease that efficiently 

digests DNA and RNA. 

dNMPs or NMPs 

dNMPs or NMPs 

and oligos with 

5´-phosphate and 

3´-OH 

di-, tri-, and 

tetra-nucleotides 

Removal of 5’-phosphorylated DNA or primers 

or oligos. Enrichment of ssDNA or ssRNA 

molecules lacking 5’-phosphate groups. 

Converting 5´ or 3´ overhangs to blunt ends. 

Removing hairpins after cDNA synthesis. 

Detecting SNPs by digesting single-base 

mismatches. With exo III, for making nested 

deletions.“S1-type” mapping of RNA. 

Removal of DNA and RNA from protein preps. 

Removal of host DNA from phage preps. 

Heat 

Inactivate 

65°C for 

10 min 

No 

No 

Nucleases 

Terminator 5´-Phosphate-Dependent Exonuclease 

Terminator Exonuclease is a 5´-to-3´, processive 

exonuclease that degrades RNAs that have a 

5´-monophosphate. RNAs with a 5-triphoaphate, 

5´-cap structure such as found on most 

eukaryotic mRNAs or a 5´-OH are resistant to 

Terminator Exonuclease degradation. It will also 

digest DNA with a 5´-monophosphate. It is not 

inhibited by proteinaceous RNase inhibitors. 

Applications 

• Characterize the 5-terminii of RNA transcripts. 

• Prepare mRNA-enriched samples from 

eukaryotic or prokaryotic total RNA 

preparations in 1 hour without the use of 

Oligo(dT), resins or magnetic beads. 


Terminator 5´-Phosphate-Dependent 

Exonuclease 

TER51020 1 U/µl 20 U 

*Patent Pending. 

Terminator 

Exonuclease 

1 Hour 

Large 

rRNA 

mRNA 

FIG 2. A 1-hour Terminator Exonuclease 

reaction digests the large rRNAs in a eukaryotic 

or prokaryotic total RNA sample, producing an 

enriched-mRNA preparation. 

FIG 3. Denaturing agarose gel analysis of E. coli 

total RNA before (-) and after (+) Terminator 

Exonuclease digestion. The Terminator Exonucleasetreated 

RNA was concentrated 10-fold. 

FIG 4. Normal rat kidney (NRK) total RNA before (A) and after (B) Terminator Exonuclease treatment. The 

Terminator Exonuclease-treated RNA was concentrated 10-fold. 


9

Mung Bean Nuclease 

Nucleases 


Mung Bean Nuclease 

M8202K 50 U/µl 2,000 U 

M8205K 50 U/µl 5,000 U 

This single-strand-specific nuclease has higher 

specificity for single-stranded DNA and RNA 

than S1 Nuclease. Unlike S1 Nuclease, Mung 

Bean Nuclease will not cleave the intact strand 

of nicked duplex DNA. 

Applications 

• Removal of hairpin structures during cDNA 

synthesis. 

• High resolution mapping of the termini and 

exon structures of RNA transcripts (commonly 

termed Berk-Sharp or S1-Mapping) using 

either internal-labeled or end-labeled probes. 

• Restriction site modification or removal by 

digestion of single-stranded protruding ends. 

• Cleavage of single-basepair mismatches. 

OmniCleave Endonuclease 


OmniCleave Endonuclease 

OC7810K 200 U/µl 10,000 U 

OC7850K 200 U/µl 50,000 U 

Provided with Dilution Buffer. 

Digests all forms of DNA and RNA including 

single-stranded and double-stranded linear, 

circular, and supercoiled. OmniCleave 

Endonuclease has the same substrate specificity 

and yields the same products as Benzonase ® , 

an enzyme derived from Serratia marcescens. 

Applications 

• Improves handling and yield of protein 

preparations by reducing the viscosity of cell 

lysates due to nucleic acids. 

• Removes trace contamination by nucleic 

acids in protein preparations. 

• Removes host DNA from phage preparations. 

Plasmid-Safe ATP-Dependent DNase 



E3101K 10 U/µl 1,000 U 

E3105K 10 U/µl 5,000 U 

E3110K 10 U/µl 10,000 U 


selectively removes contaminating bacterial 

chromosomal DNA from cosmid, BAC, fosmid, 

and plasmid preparations. The enzyme will 

processively degrade linear DNA from the 

ends; closed circular DNA (i.e. a plasmid) 

does not have free ends, and is therefore not 

degraded. These properties make Plasmid- 

Safe ATP-Dependent DNase ideal for BAC and 

fosmid purification protocols such as shot-gun 

sequencing and FISH where high purity DNA is 

a must. 

Applications 

• Removal of contaminating bacterial 

chromosomal DNA in large-scale plasmid, 

cosmid, fosmid, BAC or vector preparations. 

FIG 1. Plasmid-Safe ATP- 

Dependent DNase removes 

contaminating genomic DNA from 

plasmid preps. –, mixture of 3 µg of 

digested bacterial chromosomal DNA 

and 500 ng of uncut plasmid before 

Plasmid-Safe DNase treatment; 

+, mixture of chromosomal DNA and 

plasmid DNA after Plasmid-Safe 

DNase treatment (incubated with 

Plasmid-Safe DNase for 30 minutes 

at 37°C); M, kb ladder. 

DNA Endonucleases 

Properties of DNA Endonucleases 


Heat 

Inactivate 

Baseline-ZERO 

DNase 

dsDNA 

and 

ssDNA 

Digests dsDNA or ssDNA down to 

mononucleotides 

mononucleotides 

Removing DNA from RNA 

prep 

75°C for 

20 min 

DNase I (bovine 

pancreas) 

E.C. 3.1.21.1 

dsDNA 

and 

ssDNA 

Activated by divalent cations. In presence of 

Mg 2+ , it cleaves each DNA strand randomly 

and independently, preferentially adjacent to 

pyrimidines. In presence of Mn 2+ , it cleaves both 

strands simultaneously, generating fragments 

with blunt ends or 1-2-base overhangs. 

Oligos and dNMPs 

with 5´-phosphate 

and 3´-OH. 

Removing DNA from RNA 

preps. Random nicking of 

dsDNA. DNase footprinting. 

75°C for 

20 min 

Endonuclease IV 

(E. coli) 

dsDNA with 

abasic site 

Cleaves sugar-phosphate bond 5´ of an abasic 

site. 

dsDNA with singlenucleotide 

gaps. 

The cleaved ssDNA 

strand has a 3´-OH. 

DNA repair and anti-tumor 

drug research. Base Excision 

Sequence Scanning of DNA 

containing dUMPs. 

N/A 

T4 Endonuclease V 

UV-irradiated 

DNA with 

thymine 

dimers 

First cleaves N-glycosidic bond 5´ of thymine 

dimers, then cleaves sugar-phosphate bond 3´ 

of the abasic site. 

Nicked dsDNA with 

an abasic site at 

the 3´-end of the 

cut and thyminedimer 

bases at the 

5´-end of the cut. 

Research on repair of DNA 

exposed to UV light. 

N/A 

10 

Lambda 

Terminase 

dsDNA with 

cos sites 

Cleaves both strands at bacteriophage 

lambda cos sites. 

5´-ends with 

overhangs 12 

bases in length. 

Rapid sizing or restriction 

mapping of BAC, fosmid or 

cosmid clones. 

N/A 


Baseline-ZERO DNase 

Digests double- and single-stranded DNA to 

mononucleotides more effectively than the 

commonly used bovine pancreatic DNase I. 

Following treatment with Baseline-ZERO, even 

the small DNA oligonucleotides that remain after 

treatment with bovine pancreatic DNase I are 

undetectable. 

Provides a true zero baseline for RNA RT-PCR or 

microarray gene expression experiments. 

Applications 

• Removal of genomic DNA from RNA prior to 

RT-PCR or preparation of target RNA or cDNA 

for microarray analysis, esp. for exon arrays 

or full coverage expression analysis 

• Elimination of the DNA template following in 

vitro RNA synthesis with T7, T3 or SP6 Phage 

RNA Polymerases. 

• Removal of ssDNA and dsDNA from viral RNA. 

• Elimination of genomic DNA from RNA for 

microinjection and transfection experiments. 

• As a replacement of DNase I in applications 

requiring the removal of all DNA contamination. 

• Reverse transcription of RNA using a random 

primer since any contaminating DNA would 

also be a template for random-primed cDNA 

synthesis. 

FIG. Demonstration of 

the use of Baseline- 

ZERO DNase 

to remove small 

oligonucleotides during 

DNase treatment. Lane 

1, kilobase ladder; Lanes 

2-5, 160 ng of EcoR 

I-digested plasmid DNA 

incubated for 15 minutes 

at 37°C as follows: Lane 

2, untreated; Lane 3, 

incubated with DNase I: 

Lane 4, with supplier A’s 

hyper-active DNase; Lane 

5, with Baseline-ZERO 

DNase. Only Baseline- 

ZERO DNase removes 

the small residual oligos at 

the bottom of the gel. 

1 2 3 4 5 

Oligos 


Baseline-ZERO DNase 

DB0711K 1 U/ul 1,000 MBU 

DB0715K 1 U/ul 5,000 MBU 

*Special introductory prices 

*Patent Pending 

Nucleases 

RNase-Free DNase I 

RNase-Free DNase I (bovine pancreas) is an 

endonuclease useful in removing DNA that might 

interfere with the characterization, manipulation, 

or use of RNA, or for any application requiring 

highly purified DNase I. It efficiently hydrolyzes 

double- and single-stranded DNA to a mixture of 

short oligo- and mononucleotides. 

Applications 

• Elimination of template DNA following in vitro 

synthesis of RNA with T7, SP6, or T3 phage 

RNA polymerase. 

• Labeling of DNA by nick translation, in 

combination with Klenow or other DNA 

polymerases. 

• Treatment of RNA prior to RT-PCR. 

• Characterization of DNA-protein interactions 

by DNase I footprinting. 

1 2 3 4 

—DNA 

—transcript 


RNase-Free DNase I 

D9902K - 2,500 MBU 

D9905K - 5,000 MBU 

Supplied at a concentration of 1 U/µl. 

FIG 1. Complete DNA removal from in vitro 

transcription reactions using RNase-Free DNase I. A 

linearized DNA template was transcribed using T7 RNA 

polymerase according to standard in vitro transcription 

conditions. Lane 1, kb ladder; Lane 2, DNA control; 

Lane 3, transcription mixture; Lane 4, transcription 

mixture treated with 1 MBU of RNase-Free DNaseI for 

15 minutes at 37°C. 

Endonuclease IV, E. coli 

Endonuclease IV, cloned from the E. coli nfo 

gene, is a metalloenzyme that functions in 

vivo to repair free radical damage in DNA. The 

enzyme also has Class II abasic endonuclease 

activity, which has utility in many areas of DNA 

damage and repair research. It is also useful in 

the study of the effects of anti-tumor drugs such 

as bleomycin on nucleic acids in vivo. 

Applications 

• DNA repair research. 

• Anti-tumor drug evaluation. 


Endonuclease IV, E. coli 

E70100 2 U/µl 100 U 


T4 Endonuclease V has N-glycosylase and 

apurinic/apyrimidinic lyase (AP lyase) activities. 

Ultraviolet (UV) light produces covalent 

photoproducts in DNA, the most prevalent being 

a cis-syn cyclobutane pyrimidine dimer. T4 

Endonuclease V locates and binds to pyrimidine 

dimers in double-stranded DNA, then cleaves 

the N-glycosylic bond of the 5´-pyrimidine of 

the dimer (pyrimidine dimer DNA glycosylase) 

and breaks the phosphodiester bond 3´ to the 

resulting abasic site (3´ AP lyase). 

Applications 

• Study of UV damage to DNA and its repair, 

including DNA damage in single cells. 

• Detection of differential UV damage repair of 

transcribed sequences. 

• Detection of UV mutational hotspots. 



TE6605 20 U/µl 500 U 

TE661K 20 U/µl 1,000 U 

TE665K 20 U/µl 5,000 U 


11

Lambda Terminase 

Nucleases 


Lambda Terminase 

LT4450 2 U/µl 50 U 

LT44200 2 U/µl 200 U 

Includes 10X Reaction Buffer and 10 mM ATP Solution. 

This endonuclease encoded by bacteriophage 

lambda recognizes and cleaves DNA at cos 

sites, generating 5´-overhangs of 12 bases in 

length. Since the 12-base cos site sequence 

is rare, the sizes of inserts in BAC, fosmid, or 

cosmid clones can be rapidly determined. The 

clones are linearized with lambda terminase and 

separated by pulsed field gel electrophoresis. 

Lambda Terminase can also be used for 

chromosomal mapping and for generating 

restriction maps of DNA cloned into BAC, 

cosmid, or fosmid vectors. 

Applications 

• Rapid sizing of BAC, fosmid, or cosmid 

clones. 

• Generation of restriction maps of BAC, 

cosmid, or fosmid clone inserts. 

• Linearization of cos site-containing clones for 

in vitro packaging. 

• Specific cleavage of chromosomal DNA for 

physical mapping. 

FIG 1. Digestion of BAC clones with Lambda 

Terminase gives one band while digestion with Not 

I often gives multiple bands. DNA from six randomly 

chosen BAC clones were digested with Not I (Panel 

1) or Lambda Terminase (Panel 2) Lane M: Lambda 

Ladder PFG marker, Lane 1-6: BAC clones, Lane BT: 

BAC-Tracker Supercoiled Ladder. 

DNA Exonucleases 

Properties of DNA Exonucleases 


Heat 

Inactivate 

Exonuclease I ssDNA 3´→ 5´ exonuclease dNMPs 

Removal of ssDNA and 

oligonucleotides. 

80°C for 

15 min 

Exonuclease III 

(E. coli) 

dsDNA 

3´→ 5´ exonuclease that digests duplex DNA from 

the 3´-end of a nick or a blunt or 3´-recessed end; not 

active on thionucleotides. Exo III also has RNase H, 

3´-DNA phosphatase and apurinic DNA endonuclease 

activities. 

dNMPs and ssDNA 

on the opposite 

strand. Partial 

digestion produces 

dsDNA having 5´ 

extensions of ssDNA. 

Used with S1 Nuclease or 

Mung Bean Nuclease to make 

nested deletions. Preparation 

of ssDNA templates for 

sequencing. Site-directed 

mutagenesis. Preparation of 

labeled strand-specific probes. 

70°C for 

20 min 

Exonuclease VII 

ssDNA 

Exonuclease that digests in both 5´→ 3´ and 3´→ 5´ 

directions. 

dNMPs 

Removal of primers and 

single-stranded oligos. 

95ºC for 

10 min 

Lambda 

Exonuclease 

dsDNA 

5´→ 3´ exonuclease that digests dsDNA from 5´phosphorylated 

blunt or recessed ends. It has low 

activity on 5´-hydroxylated ends and is not active on 

nicked DNA. 

dNMPs and ssDNA 

on the the opposite 

strand. Partial 

digestion produces 

dsDNA having 3´ 

extensions of ssDNA. 

Preparation of ssDNA 

templates for sequencing. 

75°C for 

10 min 

RecBCD 

Nuclease 

(E. coli) 

dsDNA 

and 

ssDNA 

An ATP- and Mg2 +- dependent exonuclease that digests 

linear DNA in both 5´→ 3´ and 3´→ 5´ directions. Not 

active on nicked or closed-circular dsDNA. 

dNMPs 

Removal of linear DNA from 

circular DNA. 

N/A 

Rec J 

Exonuclease 

ssDNA 

5´→ 3´ exonuclease that digests ssDNA with a 5´phosphate 

or a 5´-OH. 

dNMPs 

Removal of primers and 

ssDNA from dsDNA. 

65°C for 

20 min 

T5 

Exonuclease 

dsDNA 

and 

ssDNA 

5´→ 3´ exonuclease that also has single-strandspecific 

endonuclease activity in presence of 

1-10 mM Mg 2+ ions. At

Exonuclease III, E. coli 

Exonuclease III digests duplex DNA in a 3´→ 5´ 

direction from a nick or a blunt or 3´-recessed 

end, producing stretches of single-stranded DNA 

on the opposite strand. 

Applications 

• Production of intermediates for site-directed 

mutagenesis protocols. 

• Production of strand-specific radiolabeled 

probes. 


Exonuclease III, E. coli 

EX4405K 200 U/µl 5,000 U 

EX4425K 200 U/µl 25,000 U 

Nucleases 


Exonuclease VII has high enzymatic specificity 

for single-stranded DNA and exhibits both 

5´→ 3´ and 3´→ 5´ exonuclease activities. 

It is especially useful for rapid removal of 

single-stranded oligonucleotide primers from a 

completed PCR reaction when different primers 

are required for subsequent PCR reactions. 

Exonuclease VII digestion of single-stranded 

DNA occurs in the absence of magnesium. 

Applications 

• Removal of single-stranded oligonucleotide 

primers after PCR. 

• Minimize the effect of primers left over from 

previous PCR reactions. 



EN510100 10 U/µl 100 U 

EN510250 10 U/µl 250 U 

Lambda Exonuclease 

This highly processive 5´→ 3´ 

exodeoxyribonuclease selectively digests the 

phosphorylated strand of double-stranded 

DNA. The preferred substrate is blunt-ended, 

5´-phosphorylated double-stranded-DNA. The 

enzyme has reduced activity against nicked DNA 

and against single-stranded DNA and gapped 

DNA. 

Applications 

• SSCP (single-strand conformation 

polymorphism) analysis of PCR product. 

• Generate single-stranded DNA sequencing 

template from PCR product. 



LE035H 500 U 10 U/µl 

LE032K 2,500 U 10 U/µl 


(5'-phosphate-specific 5' 3' exodeoxyribonuclease) 

P 

OH 

5'OH 

OH 

OH 

P 

OH 

P 

PCR using one primer 

with a 5'-phosphate 

5'OH 

OH 

OH 

P 

5'OH 

OH 

PCR product with strand-specific 5'-phosphate 

OH 

P 

Treatment with 


5'OH 

OH 

5'OH 

OH 

Single-stranded PCR product 

dNMPs 

FIG 1. Lambda Exonuclease selectively digests the strand of a PCR product produced using a PCR primer with 

a 5´-phosphate. The resulting single-stranded PCR product can be used for SSCP analysis or sequencing. 

RecBCD Nuclease, E. coli 

This exonuclease from E. coli degrades singleand 

double-stranded DNA. Hydrolysis of the 

DNA is bi-directional from both the 3´ and 5´ 

ends and processive, producing nucleoside 

monophosphates. Magnesium is required for 

the exonuclease activity, while calcium, nickel, 

zinc, and copper inhibit exonuclease activity. 

Calcium allows double-stranded DNA unwinding 

(helicase activity) without hydrolysis. 

Applications 

• Removal of contaminating bacterial 

chromosomal DNA in plasmid, fosmid, 

cosmid, and BAC clone or vector preparation. 


RecBCD Nuclease, E. coli 

BCD0401K - 1,000 U 


13

Rec J Exonuclease 

Nucleases 


Rec J Exonuclease 

RJ411050 10 U/µl 50 U 

RJ411250 10 U/µl 250 U 

Rec J Exonuclease, derived from E. coli, 

catalyzes removal of deoxyribonucleoside 

monophosphates from single-stranded DNA 

in a 5´→ 3´ direction. Its activity is dependent 

on Mg +2 . Rec J Exonuclease can be heatinactivated 

by incubation at 65°C for 20 

minutes. 

Applications 

• Removes primers from completed PCR 

reactions. 

• Degrades single-stranded linear DNA in 

dsDNA and plasmid preps. 

T5 Exonuclease 


T5 Exonuclease 

T5E4111K 10 U/µl 1,000 U 

This highly efficient 5´→ 3´ exonuclease for 

either single-stranded or duplex DNA has 

a tightly associated single-strand-specific 

endonuclease activity when used in the 

presence of 1-10 mM divalent magnesium ions. 

This activity may be selectively suppressed by 

using low concentrations of magnesium ions 

(< 1 mM), allowing nicked, double-stranded 

circular DNA to be “gapped” to a singlestranded 

circular species. In the absence of 

divalent metal cofactors, T5 Exonuclease is 

able to bind to DNA with a single-strand arm 

adjacent to a duplex DNA region. 

Applications 

• Plasmid mutagenesis methods. 

• Oligonucleotide site-directed mutagenesis. 

• Generation of plasmid-sequencing templates. 

• Removal of denatured DNA from alkalinebased 

plasmid purification procedures for 

improved cloning procedures. 

RNase R 

Our Exclusive RNA Exonuclease 


RNase R 

RNR07250 - 250 U 

Ribonuclease R is a magnesium-dependent 3´→ 

5´ exoribonuclease that digests essentially all 

linear RNAs but will not digest lariat or circular 

RNA structures. Intron RNA can be isolated from 

total RNA samples by digestion with RNase R. 

After digestion, only lariat structures that are 

produced during pre-mRNA splicing of intron 

regions remain. 

Applications 

• Alternative splicing studies. 

• Gene expression studies. 

• Intron cDNA production. 

• Intronic screening of cDNA libraries. 

Ribonuclease R (RNase R) 

Pre-mRNA 

Exon 1 Intron Exon 2 

Intron 

Exon 1 Exon 2 

mRNA 

Exon 1 Exon 2 

Intron 

RNase R Digestion 

Intron 

Lariat Only Remains 

i2220706 

FIG 1. How RNase R works. 

14 


RNA Endonucleases 

Properties of RNA Endonucleases 


RNase A 

RNase I, E. coli 

RNase III, E. coli 

RNase H, E. coli 

Hybridase 

Thermostable 

RNase H 

RNase T1, 

Aspergillis 

oryzae 

RiboShredder 

RNase Blend 

ssRNA 

ssRNA 

dsRNA 

RNA in 

RNA:DNA 

hybrid 

RNA in 

RNA:DNA 

hybrid 

ssRNA 

Cleaves ssRNA 3´ of pyrimidine 

residues. 

Cleaves ssRNA between all 

dinucleotide pairs. 

Cleaves dsRNA in presence of Mg 2+ 

to 12-15-bp dsRNA. Cleaves dsRNA 

in presence of 20 mM Co 2+ or Mn 2+ 

to 18-25-bp dsRNA. 

Cleaves RNA in RNA:DNA hybrid 

without affecting unhybridized RNA 

or DNA. 

Cleaves RNA in RNA:DNA hybrid 

without affecting unhybridized RNA 

or DNA. 

Cleaves ssRNA 3´ of GMPs. 

oligoribonucleotides 

with 3´-cytidine or 

3´-uridine residues 

NMPs with 5´-OH 

and 2’,3´-cyclic 

monophosphate 

dsRNA with 5´phosphate 

and 

2-base 3´-overhangs 

with 3´-OH 



and 3´-OH 



and 3´-OH 


with 3´-GMP residues 

ssRNA Efficiently degrades all RNA. NMPs 

Removal of RNA from DNA preps. RNase 

protection assays. RNA mapping and 

structure studies. 

Removal of RNA from DNA preps. RNase 

protection assays. Mismatch detection of 

single basepairs in RNA:RNA or RNA:DNA 

hybrids. 

Random cleavage of long dsRNA to short 

dsRNA. RNA interference (RNAi). 

Studies on RNA structure, RNA 

processing and maturation. 

Elimination of RNA prior to second strand 

cDNA synthesis. Removal of poly(A) tails 

from mRNA hybridized to oligo(dT). 

High-stringency hybrid selection. Highstringency 

mapping of mRNA structure. 

Transcription-based amplification methods. 

RNA mapping and structure studies. 

Removal of RNA from DNA preps. 

Removal of all RNA from genomic and 

cloned DNA preps. 

Heat 

Inactivate 

N/A 

70°C for 

15 min 

No 

5´phosphate 

65°C for 

10 min 

No 

N/A 

No 

Nucleases 

RNase A 

RNase A is an endoribonuclease that cleaves 

single-stranded RNA at the 3´-end of pyrimidine 

residues, forming oligoribonucleotides having 3´terminal 

pyrimidine-3´-phosphates. Pyrimidine- 

3´-monophosphates are also released by RNase 

A cleavage of adjacent pyrimidine nucleotides. 

Modified RNA containing pyrimidine-2’-fluorodNMPs, 

such as modified RNA made by in vitro 

transcription using EPICENTRE’s DuraScribe® 

T7 & SP6 Transcription Kits is completely 

resistant to cleavage by RNase A. 

Applications 

• Removal of RNA from DNA preparations. 

• Removal of unhybridized regions of RNA from 

DNA-RNA or RNA-RNA hybrids. 


Ribonuclease A 

MRNA092 - 

2 ml @ 

5 mg/ml 

RNase I degrades single-stranded RNA to 

nucleoside 3´-monophosphates via 2’, 3´ cyclic 

monophosphate intermediates by cleaving 

between all dinucleotide pairs. This enzyme is 

completely inactivated by heating at 70°C for 15 

minutes, eliminating the requirement to remove the 

enzyme prior to many subsequent procedures. 

Applications 


• RNase protection assays to detect singlebasepair 

mismatches in RNA:RNA and 

RNA:DNA hybrids. 



N6901K 10 U/µl 1,000 U 

N6905K 10 U/µl 5,000 U 



This endoribonuclease specifically digests 

double-stranded RNA (dsRNA) to dsRNA 

fragments that have 2-base, 3´-overhangs. 

Complete digestion of dsRNA results in dsRNA 

fragments of 12-15 bp. 

Applications 

• Digest long dsRNA to short dsRNA. 

• RNA structure studies. 

• RNA processing and maturation studies. 



RN02950 1 U/µl 50 U 



15


Nucleases 



R52250 10 U/µl 250 U 

R0601K 10 U/µl 1,000 U 

This endonuclease specifically degrades the 

RNA in an RNA:DNA hybrid, without affecting 

DNA or unhybridized RNA. It will not degrade 

double-stranded DNA or single-stranded DNA 

or RNA. E. coli RNase H is inactivated at 55°C in 

20 minutes. 

Applications 

• Elimination of RNA prior to second-strand 

synthesis of cDNA. 

• Removal of poly(A) tails from messenger RNA 

hybridized to oligo(dT). 

• Specific cleavage of mRNAs after 

hybridization to oligonucleotide probes. 

• Specific destruction of “hybrid-arrested” 

mRNAs during in vitro translation. 

• Diagnostic assays using NASBA 

transcription-based amplification methods. 

• Diagnostic assays based on the Cycling Probe 

Technology. 

• Template–dependent probe technologies. 

Hybridase Thermostable RNase H* 


Hybridase Thermostable RNase H 

H39100 - 100 U 

H39500 - 500 U 

EPICENTRE’s patented Hybridase 

Thermostable RNase H specifically degrades 

the RNA in a DNA:RNA hybrid, without affecting 

DNA or unhybridized RNA. It has optimal activity 

above 65°C and can be used at temperatures 

up to 95°C. The thermostability of the enzyme 

permits it to be used at temperatures that give 

the highest hybridization stringency for specific 

DNA:RNA heteroduplexes, maximizing sensitivity 

and selectivity while minimizing background due 

to nonspecific hybridization. 

Applications 

• High stringency hybrid selection. 

• Diagnostic assay of specific target DNA 

sequences by isothermal probe amplification. 

• Transcription-based amplification methods 

(e.g., NASBA). 

• High stringency mapping of mRNA structure. 

• Applications that require specific hydrolysis of 

the RNA in a DNA:RNA hybrid. 

*Hybridase Thermostable RNase H is covered by U.S. 

Patent Nos. 5,268,289; 5,459,055; and 5,500,370 assigned 

to EPICENTRE. This product is accompanied by a limited 

non-exclusive license for the purchaser to use the purchased 

product solely for life science research. Contact EPICENTRE for 

information on licenses to other uses. 

RNase T1, Aspergillus oryzae 


RNase T1, Aspergillus oryzae 

NT09100K - 100,000 U 

NT09500K - 500,000 U 

This endoribonuclease specifically cuts RNA 

or deaminated RNA at the 3´-end of guanosine 

residues and adjacent nucleotides through a 2’, 

3´-cyclic phosphate intermediate mechanism. 

Applications 

• RNA mapping and structure studies. 


• RNA protection assays. 

RiboShredder RNase Blend 


RiboShredder RNase Blend 

RS12100 - 100 U 

RS12500 - 500 U 

RiboShredder is a cocktail of potent RNases 

that completely degrades unwanted RNA in 

DNA purification procedures. This highly active 

cocktail contains a proprietary optimized 

blend of non-mammalian RNase enzymes. 

RiboShredder RNase Blend degrades all RNA, 

converting RNA to nucleoside monophosphates. 

• Completely degrades RNA rapidly. 

• DNase-free. 

Applications 

• Removal of RNA from genomic and cloned 

DNA preparations. 

FIG 1. Comparison of RNA-degrading capability of 

RiboShredder RNase Blend versus commonlyused 

individual RNases or other RNase cocktails. 

Nucleic acids from a standard alkaline lysis plasmid 

preparation were treated as follows under standard 

reaction conditions: Lane 1, RNase A; Lane 2, RNase I; 

Lane 3, RNase T1, Lane 4, RiboShredder RNase Blend; 

Lane 5, RNase A/RNase T1 cocktail; Lane 6, Untreated 

Alkaline lysis plasmid prep; Lane 7, supercoiled DNA 

ladder. 

16 


Base-Specific DNA Excision Mixes and DNA Glycosylases 

Properties of DNA Glycosylases and Excision Mixes 


8-Oxoguanine-DNA 

Excision Mix 

(8-Oxo-G-DNA 

glycosylase or Fpg 

and Endonuclease IV) 

Uracil-DNA Excision 

Mix 

(HK-UNG and 

Endonuclease IV) 

HK-UNG 

Thermolabile Uracil- 

N-Glycosylase (UNG) 

(UNG is also 

called Uracil-DNA 

Glycosylase or UDG) 

dsDNA 

or ssDNA 

containing 

8-oxo-GMP 

dsDNA 

or ssDNA 

containing 

dUMP 

dsDNA 

or ssDNA 

containing 

dUMP 

Cleaves sugar-phosphate 

bond 5´ of 8-oxo-GMPs. 

Cleaves sugar-phosphate 

bond 5´ of dUMPs. 

Removes uracil base 

from dUMP residues in 

DNA. 

dsDNA with single-nucleotide gaps 

where 8-oxo-G residues have been 

removed, or ssDNA strands with 

lengths equal to distances between 

8-oxo-G residues. 

dsDNA with single-nucleotide gaps 

where dUMP residues have been 

removed, or ssDNA strands with 

lengths equal to distances between 

dUMP residues. 

Uracil base and abasic DNA. Abasic 

sites can subsequently be cleaved by 

AP lyases, such as Endonuclease IV, 

or by treatment with heat or alkaline 

bases. 

Site-specific cleavage of DNA 

at 8-oxo-G sites. Base Excision 

Sequence Scanning of DNA 

containing 8-oxo-GMPs. 

Fragmentation of dUMPcontaining 

DNA. Site-specific 

cleavage of DNA at dUMP 

sites. Base Excision Sequence 

Scanning of DNA containing 


Fragmentation of dUMPcontaining 

DNA. Site-specific 

cleavage of DNA at dUMP 

sites. Base Excision Sequence 

Scanning of DNA containing 


Heat 

Inactivate 

N/A 

65°C for 

10 min 

65°C for 

10 min 

Nucleases 

8-Oxoguanine-DNA Excision Mix is a blend of 

enzymes that allows site-specific or random 

cleavage of DNA at oxidized guanine residues. 

The positions of the oxidized guanine residues 

in a DNA sequence can be mapped by sizing 

cleavage fragments on a sequencing-type gel 

from a fixed priming site, yielding data similar 

to a G-lane dideoxy-sequencing reaction. 

The enzyme mix, first depurinates oxidized 

G-residues, then cleaves the deoxyribose 

phosphate backbone at apurinic sites, 

generating a “polished” 3´-hydroxyl end and 

releasing a DNA fragment with an abasic 5´phosphorylated 

end. The minimum oligomer size 

that will serve as a substrate for excision by the 

8-Oxoguanine-DNA Excision Mix is 6 base pairs. 

The resulting DNA fragments can be analyzed 

by denaturing agarose gel or polyacrylamide gel 

electrophoresis. 

Applications 

• Mapping of G residues in any DNA. 

• DNA repair studies. 

8-Oxoguanine-DNA Excision Mix 


8-Oxoguanine-DNA Excision Mix 

OG51100 - 100 Reactions 

Contents: 8-Oxoguanine-DNA Excision Enzyme Mix, Guanine 

Oxidation Reagent, and 10X 8-OxoG-DNA Excision Reaction 

Buffer. 

Uracil-DNA Excision Mix is a blend of enzymes 

that cleave DNA at positions where uracil is 

present in place of thymine. The Uracil-DNA 

Excision Mix is useful for specific or random 

cleavage of DNA or for DNA repair studies, 

allowing mapping of uracil residues in any 

DNA. Uracil-DNA glycosylase in the Excision 

Mix removes uracil bases from DNA, creating 

a single base gap and leaving the deoxyribose 

phosphate backbone intact. Endonuclease IV in 

the Excision Mix then cleaves the DNA at each 

abasic site, leaving a 3´-hydroxyl end and an 

abasic 5´-phosphorylated end. The minimum 

oligomer size that will serve as a substrate is 6 

base pairs. Uracil-DNA Excision Mix digestion 

products can be analyzed by denaturing agarose 

gel electrophoresis or denaturing polyacrylamide 

gel electrophoresis. 

Applications 

• Mapping of uracil-containing residues in any 

DNA. 

• Mapping CpG islands. 

• DNA repair studies. 

Uracil-DNA Excision Mix 


Uracil-DNA Excision Mix 

UEM04100 - 100 Reactions 

Contents: Uracil-DNA Excision Enzyme Mix and 10X Uracil 

Excision Enzyme Buffer. 

HK-UNG Thermolabile Uracil N-Glycosylase 

This Uracil N-Glycosylase (also known as uracil- 

DNA glycosylase) hydrolyzes the N-glycosidic 

bond between the deoxyribose sugar and uracil 

in DNA containing deoxyuridine in place of 

thymidine. HK-UNG is active on both single- and 

double-stranded DNA that contains uracil, but 

has no activity on RNA or 2’-deoxyuridine-5´monophosphate. 


Fully active at 50°C; inactivated by a 10-minute 

incubation at 65°C. 

Applications 

• Repair studies of abasic sites in doublestranded 

DNA. 


HK-UNG Thermolabile Uracil N-Glycosylase 

HU59100 1 U/µl 100 U 

HU5901K 1 U/µl 1,000 U 


17

Properties of Ligases 

Ligases 

Base-Specific DNA Excision Mixes and DNA Glycosylases 

Name of Ligase Ligation Temp Cofactor 

Type of Ends 

Ligated 

Ligation Template Required to Ligate 

Blunt 

Cohesive 

Primary 

Application 

Heat 

Inactivate 

Fast-Link DNA Ligase 5-15’ @ 20°C ATP NO* YES YES Rapid Cloning 10’ @ 65°C 

T4 DNA Ligase 4°C - 25°C ATP NO* YES YES Cloning 15’ @ 65°C 

E. coli DNA Ligase 5°C - 20°C NAD YES; DNA Only Weak Activity YES Make long cDNA 20’ @ 65°C 

Ampligase ® 

DNA Ligase 

20°C - 95°C NAD YES; DNA Only NO YES 

Template- 

Dependent Ligation 

NO; Half-life 

1 hr @ 95°C 

CircLigase ssDNA Ligase 20°C - 65°C ATP NO Ligates ssDNA N/A Make ssDNA Circles 5´ @ 100°C 

T4 RNA Ligase 37°C ATP NO Ligates ssDNA Ligate RNA to DNA 

*These enzymes ligate blunt ends of dsDNA, but ligation is more efficient on a ligation template, which can be DNA or RNA. 

Make chimeric 

DNA and RNA 

15’ @ 65°C 

Fast-Link DNA Ligation Kit 


Fast-Link DNA Ligation Kit 

LK11025 - 25 Ligations 

LK0750H - 50 Ligations 

Contents: Fast-Link DNA Ligase, Fast-Link 10X Ligation 

Buffer, 10 mM ATP 

*For a full line of cloning products, please visit 

www.EpiBio.com/cloning.asp. 

This kit uses a high-quality ligase, called 

Fast-Link DNA Ligase, that was cloned at 

EPICENTRE and then formulated to provide 

extremely rapid high-efficiency DNA ligation. 

Cohesive-end ligations can be performed in 5 

minutes at room temperature. It can be used for 

routine and high-throughput DNA cloning. 

• Cohesive-end ligations in 5 minutes at room 

temperature. 

• Blunt-end ligations in 15 minutes at room 

temperature 

• Ligation of PCR product with A-overhangs in 

1 hour at 16°C. 

• Desalting of ligation products is not needed 

prior to transformation. 

• High efficiency in-gel ligation. 

Applications 

• TA cloning. 

• PCR blunt-end cloning. 

• Genomic DNA cloning and subcloning. 

• BAC library construction. 

• cDNA cloning. 

• Linker ligation. 

T4 DNA Ligase, Cloned 


T4 DNA Ligase, Cloned 

L0805H 2 U/µl 500 U 

L0810H 2 U/µl 1,000 U 

LH805H 10 U/µl 500 U 

LH810H 10 U/µl 1,000 U 

Includes 10X Reaction Buffer and a separate 25 mM ATP 

Solution. 

*For a full line of cloning products, please visit www.EpiBio. 

com/cloning.asp. 

T4 DNA Ligase is a commonly used ATPdependent 

ligase for DNA cloning. It covalently 

joins double-stranded DNA molecules having 

5´-phosphorylated and 3´-hydroxylated blunt or 

compatible cohesive ends produced by restriction 

enzyme digestion or other enzymatic processes. 

T4 DNA Ligase has no activity on single-stranded 

nucleic acids. Following a ligation reaction, T4 

DNA Ligase may be inactivated by incubation 

at 65°C for 10 minutes. EPICENTRE’s T4 DNA 

Ligase is the highest quality T4 DNA Ligase 

commercially available. 

Applications 

• Ligation of blunt or cohesive-ended DNA 

fragments. 

• Repair of nicks in double-stranded nucleic acids. 

E. coli DNA Ligase 


E. coli DNA Ligase 

DL04082H 10U/µl 200 U 

This NAD + -dependent enzyme catalyzes the 

formation of phosphodiester bonds between 

complementary 3´-hydroxyl and 5´-phosphoryl 

termini of double-stranded DNA. The enzyme 

works best with cohesive dsDNA ends and is also 

active on nicked DNA. Blunt ends can be ligated 

in the presence of condensing reagents such as 

polyethylene glycol or Ficoll. It is not effective for 

formation of DNA-RNA or RNA-RNA hybrids. 

Applications 

• Molecular cloning of dsDNA with cohesive 

ends. 

• Blunt-end ligation in presence of 10-15% 

PEG and high concentrations of monovalent 

cations. 

• cDNA cloning of products from second strand 

cDNA synthesis experiments. 

T4 RNA Ligase 


T4 RNA Ligase 

LR5010 5 U/µl 1,000 U 

LR5025 5 U/µl 2,500 U 

Includes 10X Reaction Buffer and a 10 mM ATP Solution. 

T4 RNA Ligase catalyzes the formation of a 

phosphodiester bond between a 5´-phosphorylterminated 

nucleic acid donor and a 3´-hydroxyl 

nucleic acid. The enzyme is active RNA, DNA, 

oligoribo and oligodeoxyribonucleotides, and 

nucleotide derivatives. 

Applications 

• mRNA tagging to map and sequence 5´termini 

or as a step in cDNA synthesis (RACE). 

• 3´-End labeling of RNA species. 

• Ligation of RNA and DNA species to form 

circles, extended oligonucleotides, or RNA- 

DNA-containing oligonucleotides. 

• Modifications or mutagenesis of RNA species. 

18 


X 

X 

X 

X 

Ampligase ® Thermostable DNA Ligase 

Derived from a thermophilic bacterium, stable 

and active at much higher temperatures 

than conventional DNA ligases, this enzyme 

catalyzes NAD-dependent ligation of adjacent 

3´-hydroxylated and 5´-phosphorylated termini 

in duplex DNA structures that are stable at high 

temperatures. Its half-life is 48 hours at 65°C 

and greater than 1 hour at 95°C. It has been 

shown to be active for at least 500 thermal 

cycles (94°C/80°C) or 16 hours of cycling, which 

permits extremely high hybridization stringency 

and ligation specificity. No detectable activity 

in ligating blunt-ended DNA, RNA or RNA:DNA 

hybrids. 

High thermostability allows ligation using highstringency 

hybridization conditions. 

High specificity and stringency permits sensitive 

detection of SNPs. 

Applications 

• Ligation Amplification 

(Ligase Chain Reaction, 

LCR): Ligation 

Amplification can 

distinguish between 

DNA sequences that 

differ by as little as 

a single base-pair 

and is a useful tool 

for detection of 

single nucleotide 

polymorphisms (SNPs). 

• Repeat Expansion 

Detection (RED): RED 

is a ligation-based 

method of genetic 

screening that 

detects DNA regions 

comprised of multiple 

nucleotide repeats. 

Ampligase DNA Ligase 

is used in a two-step 

thermal cycling 

reaction that generates 

oligonucleotide multimers when nucleotide 

repeats are present in a DNA template. 

• Simultaneous mutagenesis of multiple sites: 

Ampligase DNA Ligase can introduce single 

or multiple point mutations at specific sites 

by ordered ligation of PCR-amplified DNA 

fragments that have had point mutations 

introduced via mutant primers. 

• Other ligation-based detection methods. 


Ampligase® DNA Ligase Kit 

A8101 5 U/µl 1,000 U 

A30201 5 U/µl 5,000 U 

One unit of Ampligase is equal to as many as 15 units of 

other thermostable DNA ligases. Please compare competitive 

unit definitions. Contains Ampligase® DNA Ligase, 

Ampligase® 10X Reaction Buffer, and Ligation Control DNA. 

Ampligase® Enzyme and Buffer 

A0102K 100 U/µl 2,500 U 

A32250 5 U/µl 250 U 

A3202K 5 U/µl 2,500 U 



unit definitions. 25 µl of Ampligase® 10X Reaction Buffer is 

supplied with each 50 units of Ampligase® DNA Ligase. 

Ampligase® DNA Ligase 

A0110K 100 U/µl 10,000 U 

A0125K 100 U/µl 25,000 U 

A3210K 5 U/µl 10,000 U 

A3225K 5 U/µl 25,000 U 



unit definitions. Supplied as enzyme only; Reaction Buffer 

is not included. 

Ampligase® 10X Reaction Buffer 

A1905B - 5 ml 

Ampligase® 1X Storage Buffer 

A3201S - 1 ml 

Ligases 

FIG 1. Schematic of mutation discovery and 

screening using ligation amplification. The 

existence of a point mutation at the site of ligation 

interferes with oligonucleotide ligation, resulting in no 

ligation product. The lack of an amplification product 

indicates the presence of a point mutation at the 

ligation site. Oligos can also be designed so ligation 

occurs in the presence of the mutant template. 

CircLigase ssDNA Ligase 

This thermostable ATP-dependent ligase 

catalyzes intramolecular ligation (i.e., 

circularization) of single-stranded DNA (ssDNA) 

templates having a 5´-phosphate and a 3´hydroxyl 

group and ligates ends of ssDNA in 

the absence of a complementary sequence. It 

is therefore useful for making circular ssDNA 

molecules from linear ssDNA. Circular ssDNA 

molecules can be used as substrates for rolling 

circle replication or rolling circle transcription. 

Efficient single-stranded DNA ligase activity. 

Circularizes single-stranded DNA of >30 bases. 

Standard reaction conditions produce no 

detectable single-stranded DNA concatamers or 

concatameric DNA circles. 

Applications 

• Production of single-stranded DNA templates 

for rolling circle replication or rolling circle 

transcription experiments. 

• Production of single-stranded DNA templates 

for RNA polymerase and RNA polymerase 

inhibitor assays. 

FIG 1. CircLigase 

ssDNA Ligase 

converts linear 

ssDNA to circular 

ssDNA. A 71-base 

ssDNA oligo 

was converted 

to a circular 

DNA form in a 

reaction containing 

CircLigase ssDNA 

Ligase and ATP. 

Lane M, DNA markers. Lane 1, 71-base ssDNA. Lane 

2, circularization proceeds through an adenylated 

intermediate. Lane 3, the closed circular nature of 

the reaction product was confirmed by treating the 

reaction with exonuclease I, which specifically digests 

linear DNA. 


CircLigase ssDNA Ligase 

CL4111K - 1,000 U 

CL4115K - 5,000 U 

Contents: CircLigase ssDNA Ligase, CircLigase 10X 

Reaction Buffer, ATP, 50 mM MnCl2 CircLigase Linear 

ssDNA Control Substratem, Water 

*Circligase ssDNA Ligase is covered by intellectual 

property rights licensed to EPICENTRE. The purchase of this 

product conveys to the buyer the non-transferable right to 

use the purchased product and components of the product 

in research conducted by the buyer. The buyer cannot sell or 

otherwise transfer this product or its components to a third 

party and in particular, no rights are conveyed to the buyer 

to use the product or its components for commercial use 

purpose other than for research to gain information that is 

used by the buyer. 


19

Phosphatases and Kinase 

Heat-Labile Alkaline Phosphatases 

Phosphatase 

APex 

Reaction 

Temp 

20° - 50°C 

Optimal 

APex Heat-Labile Alkaline Phosphatase 


APex Heat-Labile Alkaline Phosphatase 

AP49010 1 Reaction/µl 10 Reactions 

AP49050 1 Reaction/µl 50 Reactions 

Reaction Time 

Properties of EPICENTRE’S Phosphatases 

Type of Ends Dephosphorylated 

5´- 

Protruding 

Blunt 

5´- 

Recessed 

This is a new, innovative enzyme preparation 

with improved performance over other alkaline 

phosphatases. APex Phosphatase removes 

the 5´-phosphate from all types of DNA ends, 

including 5´ protruding, blunt, and 5´ recessed 

ends, and from RNA ends. The enzyme is 

irreversibly heat-inactivated by incubation at 

70°C for 5 minutes. 

• Fast, complete and irreversible heatinactivation 

for easy transition to next step; 

no time-consuming substrate purification 

with phenol:chloroform extraction. 

• Flexible and easy to use—add directly to 

most RE buffers without supplementation; 

Nucleotides 

are Substrates 

Active at pH 

Heat 

Inactivate 

@ 37°C YES YES YES YES 5.5-12 5´ @ 70°C 

NTPhos 37°C Varies with Substrate Amt - - - YES ~7-10 15’ @ 65°C 

active over a wide range of temperatures, pH, 

salts and buffers. 

• Active on blunt, 5´- and 3´-overhang 

restricted DNA ends for compatibility with any 

restriction enzyme or experimental design. 

• One simple protocol for most applications. 

Applications 

• Dephosphorylation of DNA vectors prior to 

cloning to prevent recircularization. 

• Preparation of 5´-nucleic acid termini for 5´end 

labeling with polynucleotide kinase. 

• Dephosphorylation of DNA/RNA substrates for 

other purposes. 

Tobacco Acid Pyrophosphatase 


Tobacco Acid Pyrophosphatase (TAP) 

T19050 - 50 U 

T19250 - 250 U 

The 5´-termini of many natural RNA molecules, 

including most eukaryotic messenger RNAs, 

viral RNAs, many small nuclear RNAs, and 

heterogeneous nuclear RNAs, have a structure 

called a “cap.” Tobacco Acid Pyrophosphatase 

(TAP) hydrolyzes the phosphoric acid anhydride 

bonds in the triphosphate bridge of the cap 

structure, releasing the cap nucleoside and 

generating a 5´-phosphorylated terminus on 

the RNA molecule. The resulting “decapped” 

5´-phosphorylated terminus may be ligated to a 

3´-hydroxylated terminus using T4 RNA Ligase 

or dephosphorylated using APex Heat-Labile 

Alkaline Phosphatase for end labeling. Similarly, 

TAP digests the triphosphate group at the 5´-end 

of prokaryotic transcripts, generating an RNA 

molecule with a 5´-phosphorylated terminus. 

Applications 

• Preparation of templates for RACE (Rapid 

Amplification of cDNA Ends). 

• 5´ and 3´-end mapping of RNA. 

• Ligation of oligoribonucleotides to TAP-treated 

cellular RNA for construction of full-length 

cDNA libraries. 

• Mapping of transcription initiation sites for 

eukaryotic and prokaryotic transcripts. 

• Radiolabeling of RNA for use in sequencing or 

as a hybridization probe. 

GpppG 

FIG 1. How TAP works 

OH Capped RNA 

pG 

TAP Treatment 

OH Decapped RNA 

pG 

RNA Ligase 

T4 Polynucleotide Kinase, Cloned 


T4 Polynucleotide Kinase, Cloned 

P0505H 10 U/µl 500 U 

P0501K 10 U/µl 1,500 U 

Includes 10X Reaction Buffer without ATP. ATP is available 

separately. 

ATP Solution 

R109AT - 5 µmoles 

Provided as 500 µl of a 10 mM solution, pH 7.0. 

T4 Polynucleotide Kinase (PNK) catalyzes the 

transfer of the gamma-phosphate from ATP to 

the 5´-hydroxyl of single- and double-stranded 

DNA, RNA, and nucleoside 3´-monophosphates. 

The enzyme also removes the 3´-phosphate 

from 3´-phosphoryl polynucleotides, 

deoxyribonucleoside 3´-monophosphates, and 

deoxyribonucleoside 3´, 5´-diphosphates to form 

a 3´-hydroxyl group. 

Applications 

+ 

pG pG OH 

+ 

additional ligation 

products 

• Labeling of 5´-termini of DNA and RNA for DNA 

sequencing, blot-hybridization, or transcript 

mapping. 

• Phosphorylation of oligonucleotide linkers and 

other DNA or RNA molecules prior to ligation, 

or for use in ligation amplification with 

Ampligase ® Thermostable DNA Ligase. 

• Preparation of labeled DNA or RNA molecular 

weight markers for gel electrophoresis and 

chromatography. 

Ligated RNA 

i490407tap2 

20 


EasyLyse Bacterial Protein Extraction Solution 

This extraction solution is designed for lysing 

bacterial cells for the isolation of proteins, 

especially recombinant gene products 

expressed in E. coli, without significant loss of 

enzymatic activity. It contains a highly active 

enzyme for cell lysis and a potent nuclease that 

reduces extract viscosity by digesting all nucleic 

acids in the sample. The EasyLyse Solution is 

formulated as a homogeneous reagent for ease 

of use in high-throughput applications without 

sonication. 

Gives higher yields of soluble protein. 

Applications 

• Rapid protein screening. 

• Easy protein purification. 

• Enzymatic studies. 

• ELISA studies. 

• Manual or robotic procedures. 

Bacterial Lysis (%) 

Specific Activity of Soluble LDH (nmol/min/µg) 

100 

80 

60 

40 

20 

0 

EasyLyse Bacterial Lysis Efficiency 

EasyLyse 

Supplier P 

EasyLyse Preserves Enzyme Activity 

1.5 

1.0 

0.5 

0.0 

1.2 

EasyLyse 

0.12 

Supplier P 

c180308bnl 

c170308bnl 

FIG 1. Lysis of 

E. coli aliquots 

with soluble 

protein quantities 

expressed as a % 

of total protein, as 

determined by the 

Coomassie Plus 

Protein Assay. 

FIG 2. E. coli Lactate 

Dehydrogenase 

(LDH) specific 

activity expressed 

as nmol/min/µg of 

soluble cell protein. 


EasyLyse Bacterial Protein Extraction Solution 

500, 1-ml 

Purifications 

RP03750 - 

or 

48, 96-well 

Microplates, 

100 µl/well 

Contents: Lysis Buffer, Enzyme Mix, MgCl2 Solution. 

Lysozymes 

Ready-Lyse Lysozyme Solution for Protein Extraction 

Ready-Lyse Lysozyme Solution is a nonmammalian, 

non-avian, recombinant lysozyme 

preparation for the lysis of Gram-negative (such 

as E. coli) and Gram-positive (such as Bacillus 

sp.) bacteria. The specific activity of Ready-Lyse 

Lysozyme is 200-fold higher than the specific 

activity of egg white lysozyme. Also, unlike egg 

white lysozyme, Ready-Lyse Lysozyme Solution 

is stable at –20°C, eliminating the need to 

prepare a fresh solution for each use. The use of 

Ready-Lyse Lysozyme results in higher yields of 

protein than can be obtained with standard egg 

white lysozyme. 

Applications 

• Lysis of Gram-negative or Gram-positive 

bacteria for protein purification. 

Table 1. Bacteria lysed with Ready-Lyse Lysozyme 

Solution. 

Gram-negative 

Escherichia coli 

Salmonella typhimurium 

Actinobacillus 

pleuropneumoniae 

Rhodobacter sphaeroides 

Shewanella putrefaciens 

Flavobacteria odoratum 

Gram-positive 

Oerskovia xanthinolytica 

Bacillus subtilis 

M 

0 hr. 

1 hr. 

3 hr. 

1 2 3 4 

FIG 1. Use of Ready-Lyse Lysozyme Solution to 

recover recombinant proteins. One ml of induced 

cells from a recombinant E. coli clone was pelleted by 

microcentrifugation before induction and at 1 and 3 

hours after induction. 


Ready-Lyse Lysozyme Solution 

R1802M - 2 X 10 6 U 

R1810M - 10 X 10 6 U 

*Animal Product-Free Ready-Lyse Lysozyme Solution 

is available. 

**For a complete line of purification products, please visit 

www.EpiBio.com. 

Ready-Lyse Lysozyme Solution for Nucleic Acid Extraction 

Ready-Lyse Lysozyme Solution is a nonmammalian, 

non-avian, recombinant lysozyme 

preparation for the lysis of many Gram-negative 

bacteria such as Escherichia coli and Grampositive 

bacteria such as Bacillus subtilis. The 

use of Ready-Lyse Lysozyme results in higher 

yields of DNA and RNA than obtained with 

standard egg white lysozyme. 

Due to its higher specific activity, less Ready- 

Lyse is needed for lysis compared to egg white 

lysozyme. This reduces loss of nucleic acid from 

enzyme binding. 

Supplied as a ready-to-use solution stable at 

–20°C, so there is no need to prepare fresh 

solution prior to each use or to freeze aliquots 


that are used once and then discarded, as is 

required with egg white lysozyme. 

Applications 

• Lysis of Gram-negative and Gram-positive 

bacteria for preparations of nucleic acids. 

• In-well gel screening of recombinant DNA in 

agarose gels. 

FIG 1. Lysis with Ready-Lyse 

Lysozyme increases yields of 

nucleic acids. Approximately 

50% of the DNA was lost due to 

precipitation by egg white lysozyme 

(EW), while Ready-Lyse Lysozyme 

(RL) caused minimal precipitation 

losses of DNA compared to control 

(C) samples without lysozyme. 


Ready-Lyse Lysozyme Solution 

R1802M - 2 X 10 6 U 

R1810M - 10 X 10 6 U 

*Animal Product-Free Ready-Lyse Lysozyme Solution is 

available. 

**For a complete line of purification products, please visit 

www.EpiBio.com. 

Red Cell Lysis Solution 

MRC0912H - 1200 ml 

Tissue & Cell Lysis Solution 

MTC096H - 600 ml 

21

RecA Protein, E. coli 

Other Protein products 


RecA Protein, E. coli 

RC44200 5 µg/µl 200 µg 

RC441MG 5 µg/µl 1 mg 

This multi-functional DNA-binding protein 

encoded by E.coli plays integral roles in both 

homologous recombination and post-replicative 

DNA repair mechanisms. In vitro, RecA Protein 

helps promote homologous recombination 

through a multiple-step ATP-dependent 

pathway. Initially, the protein binds preferentially 

to single-stranded DNA forming a nucleoprotein 

filament. The filament complex binds to naked 

duplex DNA and searches for regions of 

homology. Once a region of homology is found, 

strand displacement and exchange begins. 

Applications 

• Site-directed mutagenesis through 

displacement loop structures. 

• Targeted site-specific cleavage of small and 

large DNA. 

• Enrichment of target sequences from libraries 

or other DNA pools. 

• Visualization of DNA for electron microscopy. 

• Cloning or other experiments involving 

use of RecA Protein and a site-specific 

oligonucleotide to block endonuclease 

cleavage at the complementary site in a 

target DNA molecule. 

Single-Stranded DNA Binding Protein (SSB), E. coli 


Single-Stranded DNA Binding Protein (SSB) 

SSB02200 2 mg/ml 200 µg 

DNA Topoisomerase I, Vaccinia 

Single-Stranded DNA Binding binds singlestranded 

DNA with high specificity. In vivo, SSB 

is involved in DNA replication, recombination, 

and repair. In vitro, SSB enhances several 

molecular biology applications by destabilizing 

DNA secondary structure and increasing 

the processivity of polymerases. E. coli SSB 

is also required for in vitro transcription of 

single-stranded DNA templates by MiniV 

RNA Polymerase, a transcriptionally-active 

1,106-amino acid domain of the N4 virion RNA 

polymerase. 

Applications 

• Transcription of ssDNA templates by MiniV 

RNAP. 

• Targeting restriction endonuclease digestion 

to any restriction enzyme site in cloned 

single-stranded DNA. 

• Enhance the specificity and yield of PCR 

reactions. 

• Improve DNA sequencing results through 

regions with strong secondary structure. 

• Site-directed mutagenesis when used in 

conjunction with recA protein. 

• Improve the processivity of DNA polymerases. 

• DNA replication and recombination studies. 


DNA Topoisomerase I, Vaccinia 

VT710500 10 U/µl 500 U 

VT7105K 10 U/µl 5,000 U 

Topoisomerase I from vaccinia virus is a type 

I eukaryotic topoisomerase that removes both 

positive and negative superhelical turns (also 

called right- and left-handed supercoils) from 

covalently closed DNA. The product of the 

reaction is a covalently closed, circular DNA with 

fewer positive or negative superhelical turns. 

DNA Topoisomerase I does not absolutely require 

Mg 2+ to function, although low concentrations of 

magnesium ions may increase activity. 

Applications 

• Studying the effects of supercoiling on 

transcription in vitro. 

• Studying chromatin reconstitution in vitro. 

• Determining the degree of supercoiling of 

naturally occurring DNA. 

• Detecting mutant plasmids that differ in 

length by only one basepair. 

• Increasing restriction endonuclease digestion 

of resistant DNA substrates by “unwinding” 

the DNA coils to expose restriction sites. 

22 


Tagetin RNA Polymerase Inhibitor 

Tagetin RNA Polymerase Inhibitor is the only 

compound known to potently and selectively 

inhibit RNA polymerase III from a variety of 

eukaryotic organisms including mammalian 

cells, Saccharomyces cerevisiae, Drosophila 

melanogaster, Bombyx mori, and Xenopus 

laevis oocytes. It strongly inhibits Escherichia 

coli RNA polymerase and plant chloroplast RNA 

polymerase. Plant nuclear RNA polymerases 

I, II, and III are much less sensitive to Tagetin 

Inhibitor. Phage-encoded RNA polymerases such 

as SP6 and T7 are also relatively insensitive. 

With both eukaryotic and prokaryotic RNA 

polymerases, the degree of inhibition is 

template-dependent. 

Applications 

• RNA polymerase studies. 

• Transcription studies. 

FIG 1. Tagetin Inhibitor 

activity on E. coli RNA 

Polymerase. Each gel 

lane shows products of 

a standard transcription 

reaction using a 

bacteriophage template, 

1 U of E. coli RNA 

Polymerase Holoenzyme, 

and varying amounts of Tagetin Inhibitor. Lane 1, 100 

U; Lane 2, 10 U; Lane 3, 1 U; Lane 4, 0.1 U; Lane 

5, control without Tagetin Inhibitor. 50% inhibition of 

transcription is seen at 1 U Tagetin Inhibitor per unit of 

E. coli RNA Polymerase. 


Tagetin RNA Polymerase Inhibitor 

T9705H 20 U/µl 500 U 

T9702K 20 U/µl 2,500 U 

Enzyme and Protein Inhibitors 

ScriptGuard RNase Inhibitor 

ScriptGuard RNase Inhibitor is your best 

defense against common RNases including 

RNase A, RNase B, and RNase C. This 

recombinant RNase inhibitor protein provides 

reliable protection of your precious RNA samples 

by binding strongly to RNases in a 1:1 ratio. 

EPICENTRE’s ScriptGuard RNase Inhibitor is 

free of unwanted contaminants that can plague 

other commercially available preparations of 

RNase inhibitors. 

• A potent affinity for RNases (K i 

>10 -14 M) 

ensures rapid inhibition even when trace 

amounts of RNase are present. 

• Free of detectable RNase or DNase activity 

and mammalian DNA. 

• Does not interfere with enzymes commonly 

used to prepare or analyze RNA. 

• Less sensitive to oxidation than traditional 

RNase inhibitors. 

Applications 

• Effectively inhibits the degradation of RNA by 

eukaryotic RNases in a variety of applications, 

including cDNA synthesis, RT-PCR and in vitro 

transcription and translation. 


ScriptGuard RNase Inhibitor 

SRI6325 40 U/µl 2,500 U 

SRI6310K 40 U/µl 10,000 U 

Protein Transport Inhibitor Brefeldin A 

Brefeldin A (BFA), a metabolite of the fungus 

Eupenicillium brefeldianum, specifically 

and reversibly blocks protein transport from 

the endoplasmic reticulum (ER) to the Golgi 

apparatus in many cell types and species. 

These effects are generally accompanied by 

distinct morphological changes, including the 

apparent collapse of the Golgi stacks. The fast 

and reversible redistribution of intracellular 

membranes is accompanied by various specific 

and reversible effects on cellular protein traffic, 

including protein transport from the ER to the 

Golgi, protein secretion, vesicular assembly, 

antigen presentation, trans- and endocytosis, 

and viral assembly and budding. 

Applications 

• Studying mechanisms of protein transport 

and targeting. 

• Inducing “retrograde transport” of proteins 

normally resident in the Golgi into the ER. 

• Blocking protein secretion in many cell types. 

• Blocking antigen presentation by major 

histocompatibility complex class I and class II 

molecules. 

• Reversibly arresting assembly and release of 

viral particles. 

• Blocking the toxic effects of ricin, modeccin, 

abrin, and Pseudomonas toxin in various cell 

types. 

• Mapping post-translational modifications 

of cell-surface receptors and other 

glycoproteins. 

FIG 1. Treatment 

of primary mouse 

pituitary cells with 

Brefeldin A. BFA 

treatment results in 

the redistribution of 

Golgi membranes 

into the endoplasmic 

reticulum (+BFA) as seen when compared with an 

untreated cell (-BFA). (Electron micrographs are 

courtesy of J.A. Magner, Michael Reese Hospital, 

University of Illinois, Chicago.) 


Brefeldin A 

B901MG - 1 mg 

B905MG - 5 mg (5 x 1 mg) 

Enzyme Storage Buffer 

Composition 

50% glycerol containing 50mM Tris-HCl (pH 

7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, and 

0.1% Triton X-100. 

Applications 

• Enzyme storage or dilution. 


Enzyme Storage Buffer 

ESB4901 - 1 ml 


23

Specialty Enzymes for DNA and RNA Research 

Contents 

RNA Polymerases and Replicases page 3 

RNA Capping and Tailing Enzymes page 4 

DNA Polymerases page 6 

Reverse Transcriptase page 8 

Nucleases and Glycosylases page 9 

Ligases page 18 

Phosphatases and Kinase page 20 

Lysozymes page 21 

Other Protein products page 22 

Enzyme Inhibitors page 23 

EPICENTRE has the necessary infrastructure, qualifi ed and trained personnel and Quality 

Systems procedures that are required to manufacture molecular biology enzymes of high 

purity and quality that will exceed the expectations of the customer. 

EPICENTRE develops, manufactures and sells optimal enzyme systems and reagents for 

life science research, diagnostics and pharmaceutical bioprocessing. 

LARGE QUANTITIES & SPECIAL FORMULATIONS are available. Please call toll free (U.S. 

only) 1-800-284-8474 to inquire about custom kits or enzyme formulations, highthroughput 

packaging, bulk orders, animal product-free reagents and manufacturing to 

meet specifi c regulatory requirements. 

EPICENTRE never stops introducing new enzymes for your research needs. Sign up with us 

at www.EpiBio.com/reply_card.asp to keep updated. 

USA: 800-284-8474 

Technical Support 

Tel: 608-258-3080 USA: 800-284-8474 

Fax: 608-258-3088 

techhelp@EpiBio.com

EPICENTRE Enzyme Catalog-Beta/Int'l version

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