Science Catalog 2012 Life Cell Signaling RNA Purification PureYield™ RNA Midiprep System Product PureYield™ RNA Midiprep System Available Separately RNA Lysis Buffer (RLA) RNA Wash Solution (RWA) Red Blood Cell Lysis Solution (CLB) Eluator™ Vacuum Elution Device Size Cat.# Price (Fr) 10 preps Z3740 180.00 50 preps Z3741 765.00 50 ml Z3051 72.00 58.8 ml Z3091 71.00 200 ml Z3141 82.00 4 each A1071 161.00 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. RNA purified with the PureYield™ RNA Midiprep System has no detectable genomic DNA contamination. Total RNA was isolated from 1 × 108 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 104, 103, 102 and 101 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. 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 PureYield™ RNA Midiprep System Technical Manual Part# TM279 Average Yields of Total RNA Isolated from Tissues and Cells. Maximum Sample Type Rat Tissues Liver Lung Bacteria E. coli Cell Line HEK 293T HeLa Amount to Process 300mg 300mg 1 ⋅ 1010 cells 5 ⋅ 107 cells 5 ⋅ 107 cells Average Yield per Prep (µg)1 1025.8 217.0 782.7 453.3 329.2 Average A260/A230 1.7 1.9 2.5 2.1 1.8 Average A260/A280 1.8 2.1 2.1 1.9 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 124 For complete and up-to-date product information visit: www.promega.com/catalog 5646TE Pagina 127
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