2012 Promega catalogue

Cell Signaling
TSAP Thermosensitive Alkaline Phosphatase
Product Size Cat.# Price ($)
TSAP Thermosensitive Alkaline Phosphatase 100 units M9910 113.00
For Laboratory Use.
Description: TSAP Thermosensitive Alkaline Phosphatase catalyzes the
removal of 5´ phosphate groups from DNA, thus preventing the recircularization
and religation of linearized cloning vector DNA during ligation. It is effective on
3´ overhangs, 5´ overhangs and blunt ends. It is also useful for preparing DNA
for 5´ end-labeling by removing existing phosphate groups from the 5´ end.
TSAP is irreversibly inactivated by heating at 74°C for 15 minutes.
Therefore, a DNA cleanup step is not required before proceeding to a ligation
reaction. TSAP is fully active in all restriction enzyme reaction buffers tested
under the conditions listed below, facilitating a streamlined restriction digestion,
dephosphorylation and ligation reaction.
Features:
• Easy To Use: TSAP is active in all Promega restriction enzyme buffers,
eliminating any cleanup steps or buffer swaps.
• Convenient: TSAP is irreversibly inactivated by heating at 74°C for
15 minutes. This allows streamlining of the restriction enzyme digestion,
dephosphorylation and ligation procedure by eliminating the need for
cleanup after alkaline phosphatase treatment.
• Blue/White Cloning-Qualified: Promega’s blue/white cloning assay
provides a higher level of quality control for enzymes used in cloning
applications.
• Provided with Promega MULTI-CORE Buffer.
Storage Conditions: Store at –20°C. See the expiration date on the label.
Protocol Part#
TSAP Thermosensitive Alkaline Phosphatase Protocol 9PIM991
Comparison of Alkaline Phosphatases.
TSAP CIAP*
Heat Inactivated Yes
No
Inactivation Temperature 74
N/A
Incubation Time 15 min 2 × 30 min
Special Buffer Required/Recommended
Active in all Promega
No
Yes
Restriction Enzyme Buffers Yes
No
Units required in different RE Buffers 1–2
N/A
Blue/White Cloning-Qualified Yes
Yes
Only TSAP does not require a special buffer and is active in all Promega restriction
enzyme buffers, making it the most convenient and cost-effective choice.
CIAP = Calf Intestinal Alkaline Phosphatase
For complete and up-to-date product information visit: www.promega.com/catalog
9493LA
Polymerases
DNA Polymerase I
Product Size Conc. Cat.# Price ($)
DNA Polymerase I 500 u 5–10 u/µl M2051 128.00
For Laboratory Use.
2,500 u 5–10 u/µl M2055 483.00
Description: DNA Polymerase I catalyzes the template-directed polymerization
of nucleotides into duplex DNA in a 5´→3´ direction. DNA Polymerase I
possesses a 3´→5´ exonuclease activity or “proofreading” function, which
lowers the error rate during DNA replication, and also contains a 5´→3´
exonuclease activity, which enables the enzyme to replace nucleotides in
the growing strand of DNA by nick translation. The enzyme, purified from
recombinant E. coli, is capable of catalyzing de novo synthesis of synthetic
homopolymers and provides a convenient method for the preparation of a
variety of defined DNA substrates.
Features:
• Flexible: DNA Polymerase I may be used in a variety of molecular
applications.
• May Be Heat-Inactivated: DNA Polymerase I is inactivated by heating at
68°C for 10 minutes.
• Provided with 10X Reaction Buffer: 500mM Tris-HCl (pH 7.2 at 25°C),
100mM MgSO 4, 1mM DTT.
Storage Conditions: Store at –20°C.
Protocol Part#
DNA Polymerase I Protocol 9PIM205
DNA Polymerase I Large (Klenow) Fragment
Product Size Conc. Cat.# Price ($)
DNA Polymerase I Large (Klenow) 150 u 5 u/µl M2201 117.00
Fragment
For Laboratory Use.
500 u 5 u/µl M2206 344.00
Description: DNA Polymerase I Large (Klenow) Fragment is a DNA-dependent
DNA polymerase that lacks the 5´→3´ exonuclease activity of intact E. coli
DNA Polymerase I but retains its 5´→3´ polymerase, 3´→5´ exonuclease and
strand displacement activities. The enzyme is a 68kDa C-terminal fragment of
DNA Polymerase I. The 5´→3´ polymerase activity of Klenow Fragment can be
used to fill in 5´-protruding ends with unlabeled or labeled dNTPs, to sequence
single- or double-stranded DNA templates, for in vitro mutagenesis using
synthetic oligonucleotides, for cDNA second-strand synthesis and to generate
single-stranded DNA probes. The 3´→5´ exonuclease activity can be used to
generate blunt ends from a 3´-overhang.
Features:
• Flexible: DNA Polymerase I Large (Klenow) Fragment may be used in a
variety of molecular applications. It is also active in many Promega 1X
restriction enzyme buffers.
• May Be Heat-Inactivated: DNA Polymerase I Large (Klenow) Fragment is
inactivated by heating at 75°C for 10 minutes.
• Provided with 10X Reaction Buffer: 500mM Tris-HCl (pH 7.2 at 25°C),
100mM MgSO 4, 1mM DTT.
• Choose Your Configuration: Learn more about our custom options for
this product at: www.promega.com/myway/
Storage Conditions: Store at –20°C.
Protocol Part#
DNA Polymerase I Large (Klenow) Fragment Protocol 9PIM220
89
4
Cloning and DNA Markers
Section
Contents
Table of
Contents