Cell Signaling Reverse Transcription System Product Reverse Transcription System Available Separately Magnesium Chloride Solution Reverse Transcription 10X Buffer Size Cat.# Price (Fr) 100 reactions A3500 722.00 Size Conc. Cat.# Price (Fr) 1.5 ml 25 mM A3511 60.00 1.4 ml A3561 60.00 A3511, A3561 For Laboratory Use. A3500 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The Reverse Transcription System provides reagents to efficiently reverse transcribe RNA into cDNA in fifteen minutes. The cDNA prepared from each reaction using this system may be used directly in multiple PCR amplifications using Taq DNA polymerase. The AMV Reverse Transcriptase synthesizes single-stranded cDNA from total or poly(A)+ RNA. Both Oligo(dT)15 and Random Primers are included, allowing cDNA synthesis from virtually any RNA source. The system contains sufficient reagents for 100 cDNA synthesis reactions, processing 1μg of RNA per reaction. Each cDNA synthesis reaction may be divided and used in up to 20 separate PCR amplifications. A polyadenylated 1.2kb RNA transcript is provided as a control template for the cDNA synthesis reaction. Features: • Fast: Efficiently reverse transcribe poly(A)+ mRNA or total RNA in fifteen minutes. • Convenient: PCR-compatible components are provided in optimized volumes for 100 reactions. • Positive Controls: A polyadenylated RNA transcript is provided to help troubleshoot RT-PCR parameters. Storage Conditions: Store at –20°C. Store Positive Control RNA at –70°C. Protocol Reverse Transcription System Technical Bulletin Part# TB099 AccessQuick™ RT-PCR System Product AccessQuick™ RT-PCR System Size Cat.# Price (Fr) 20 reactions A1701 200.00 100 reactions A1702 775.00 500 reactions A1703 3100.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The AccessQuick™ RT-PCR System is an easy and convenient master mix system for setting up one-tube RT-PCR. It is designed to increase the convenience of performing RT-PCR by combining these components in a single tube: Tfl DNA Polymerase, dNTPs, magnesium sulfate and reaction buffer. The AMV RT enzyme is provided in a separate tube to allow important, no-RT control reactions. The AccessQuick™ Master Mix is simply added to RNA templates in reaction vials, followed by the AMV RT, primers and water. The AccessQuick™ RT-PCR Master Mix is intended for routine RT-PCR applications that have been previously optimized and do not require extreme conditions. Features: • Achieve Maximum Convenience: Save yourself four pipetting steps. Simply combine the AccessQuick™ Master Mix, AMV RT, your genespecific primers, your RNA template and water. Separate AMV RT allows important, no-RT control reactions. • Use Less Template: Amplify from zeptomole (10–21mol) levels of RNA. • No Buffer Additions Required: Set up reactions in a single tube, drop into a thermal cycler, come back later for results—no additions between the reverse transcription and DNA amplification steps. • Stable: System components are stable over many freeze-thaw cycles. Storage Conditions: Store all system components at –20°C. Protocol AccessQuick™ RT-PCR System Protocol zeptomoles RNA per reaction 05 10 MM bp 323 – Stability of AccessQuick™ Master Mix through multiple freeze-thaw events. Rapid freeze-thaw events were performed 0, 5 and 10 times by removing a sample of the AccessQuick™ Master Mix from –70°C storage and placing it in a 50°C heat block. After 5 cycles, and again after 10 cycles, we added AMV RT, primers and RNA. All samples were then used in RT-PCR reactions to amplify a 323bp fragment using the indicated amounts of the 1.2kb Kanamycin Positive Control RNA (Cat.# C1381) template. Lane M = 100bp DNA Ladder (Cat.# G2101). Part# 9PIA170 12 For complete and up-to-date product information visit: www.promega.com/catalog 217 PCR 3511TB 0 2,500 252.50.250 2,500 252.50.250 2,500 252.5 0.25 Pagina 220
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