2012 Promega catalogue

Cell Signaling
PureYield RNA Midiprep System
Product Size Cat.# Price ($)
PureYield RNA Midiprep System 10 preps Z3740 188.00
50 preps Z3741 816.00
Available Separately
RNA Lysis Buffer (RLA) 50 ml Z3051 67.00
RNA Wash Solution (RWA) 58.8 ml Z3091 67.00
Red Blood Cell Lysis Solution (CLB) 200 ml Z3141 82.00
Eluator Vacuum Elution Device 4 each A1071 77.00
Z3740, Z3741, Z3141, A1071 For Laboratory Use. Z3051, Z3091, Z3370, Z3372, Z3374 For
Research Use Only. Not for Use in Diagnostic Procedures.
Description: The PureYield RNA Midiprep System isolates intact, pure total
RNA from essentially any sample type for use in a wide range of applications.
The use of a novel Clearing Agent enables the rapid purification of total RNA
with undetectable levels of genomic DNA contamination without using DNase.
A novel combination of reagents, membranes and protocol leads to yields of
up to 1mg of total RNA without organic solvents, protease digestions or alcohol
precipitations. One kit can be used to isolate pure total RNA from a wide variety
of sample types, such as tissues, cultured cells, bacteria, yeast, plants and
blood. The protocol also can be adapted for other sample types.
Commonly used methods provide total RNA that is contaminated with genomic
DNA. This contamination can interfere with sensitive methods, such as realtime
RT-PCR and microarray analysis. The PureYield RNA Midiprep System
avoids this problem by selectively removing the genomic DNA prior to total RNA
purification. The eluted total RNA is free of detectable DNA and ready for use in
sensitive downstream applications.
The system has been designed for use with centrifugation or vacuum (e.g., the
Vac-Man ® Laboratory Vacuum Manifold) formats.
The Eluator Vacuum Elution Device is used to elute nucleic acids from
PureYield Midiprep or Maxiprep columns. It consists of two pieces, a
blue base and a clear column assembly. The base interfaces with a Vacuum
Manifold that contains Luer-Lok ® fittings, such as the Vac-Man ® Laboratory
Vacuum Manifold (Cat.# A7231), and holds a 1.5ml tube to capture the eluted
nucleic acids. The column assembly accepts PureYield Midiprep or Maxiprep
columns.
The Eluator Device eliminates the requirement for a centrifuge with a
swinging bucket rotor for nucleic acid purification, simplifying and speeding
purification protocols.
For complete and up-to-date product information visit: www.promega.com/catalog
Features:
• Enhanced Results: Purified total RNA with undetectable genomic DNA
contamination improves results in downstream applications.
• Improved Productivity: Purifying total RNA without the use of DNase
treatment reduces steps during purification and in downstream
applications.
• Safety and Efficiency: Rapid purification of high yields of total RNA
without the use of hazardous organic solvents.
• Flexibility: Single system for purifying total RNA directly from cultured
cells, bacteria, yeast, plants and other sample types.
Storage Conditions: Store the RNA Lysis Buffer (RLA) with added
β-Mercaptoethanol (BME) at 4°C. Store all other components at 22–25°C.
Protocol Part#
PureYield RNA Midiprep System Technical Manual TM279
RNA purified with the PureYield RNA Midiprep System has no
detectable genomic DNA contamination. Total RNA was isolated from
1 × 10 8 HEK 293T cells using the PureYield RNA Midiprep System, a
competitor’s kit and a competitor’s reagent. One hundred nanograms of each
total RNA sample was assayed using the Plexor ® qPCR System (Cat.# A4011)
to detect genomic DNA contamination. Human Genomic DNA (Cat.# G3051)
in quantities of 10 4 , 10 3 , 10 2 and 10 1 copies was used as a standard. The
PureYield RNA Midiprep System samples showed no detectable genomic
DNA. Competitor Q and Competitor T showed an average of 227 and 17
copies, respectively. The horizontal purple line in the upper right corner of this
figure indicates no detectable genomic DNA in the PureYield RNA Midiprep
System sample.
Average Yields of Total RNA Isolated from Tissues and Cells.
Maximum Average Yield Average Average
Sample Type Amount to Process per Prep (µg) 1 Rat Tissues
A260/A230 A260/A280 Liver 300mg 1025.8 1.7 1.8
Lung
Bacteria
300mg 217.0 1.9 2.1
E. coli 1 × 10 10 Cell Line
cells 782.7 2.5 2.1
HEK 293T 5 × 107 cells 453.3 2.1 1.9
HeLa 5 × 107 cells 329.2 1.8 2.0
1 The average total RNA yield shown is from a 1ml elution. A second 1ml elution yielded an additional average of 366.4µg (rat liver), 47.0µg (rat lung), 196.8µg (E. coli), 45.7µg (HEK 293T cells) and
73.8µg (HeLa cells) of total RNA. 9486LA
5646TE
125
5
DNA and RNA Purification
Section
Contents
Table of
Contents