RScript Reverse Transcriptase

RScript Reverse Transcriptase
RScript Reverse Transcriptase
RScript Reverse Transcriptase

Catalog numberRT001-0050/RT001-0250

Size50 rxns/250 rxns

Concentration200 U/μl

StorageStable for up to 2 years at -20°C

$96 (50 rxns)$384 (250 rxns)
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Note: RT enzyme / MMLV RTase / RNA template / Complementary DNA / cDNA / Reverse Transcription / Molecular cloning / RNA sequencing / PCR / Genome Analysis / superscript

Description

Overcome the most challenging RNA structures over a wide temperature range.

RScript Reverse Transcriptase - Engineered innovatively and specifically for both Research and Diagnostic applications for meeting all your cDNA synthesis needs and for overcoming the most challenging secondary RNA structures over a wide temperature range. Our next-generation, engineered recombinant M-MLV reverse transcriptase, with improved thermostability, processivity, robustness, optimal cDNA yields, proprietary site mutations for reduced RNase H activity, and extended half-life, is the most versatile reverse transcriptase in the world for not only simply meeting the routine cDNA synthesis requirements but also enabling superior performance for even the most challenging RNA samples at hand.

Kit Contents

RT001-0050RScript Reverse Transcriptase 50 μl x 1 vial
2X Sharp reaction buffer 500 μl x 1 vial
RT001-0250RScript Reverse Transcriptase 250 μl x 1 vial
2X Sharp reaction buffer 500 μl x 5 vials
Features
  • Excellent reverse transcriptase for challenging COVID-19 RNA detection.
  • Enzyme activity stable even when stored at 37°C over 21 days.
  • Enhanced thermostability from 42°C to 60°C.
Primer Selection

Primer amounts recommended for efficient cDNA synthesis are 0.5 μg of oligo(dT) (anneal to the 3'-poly(A) + mRNA) or random primers (anneal at non-specific sites of RNA templates), or 2 μM of gene-specific primers per 20 μl reaction.

Reaction Setup

cDNA Synthesis

1. For each 20 μl cDNA synthesis reaction, assemble the following in a PCR tube. Keep it on ice just prior to use.

ComponentFinal conc.Volume
RNA template*10 pg-2 ug total RNA or 10 pg-500 ng mRNAX μl
2X Sharp Reaction Mix (including dNTPs,MgCl2)*10 μl
Primers1 μl
RScript Reverse Transcriptase200 U1 μl
RNase Inhibitor20-40 U1 μl
dNTPs (10 mM)0.2 mM0.4 μl
Nuclease-Free WaterAdd to 20 μl
Total volume20 μl

*RNA template, primers and 2X Sharp reaction buffer need to be premixed and heated at 65°C for 5 minutes in advance. After RNA has been pre-heated and incubated on ice bath at least 1 minute, add other components according to the table.

2. Mix the reaction solution gently by pipetting.

3. Cap the tubes and place them in the temperature-controlled water bath or heat blocks. Incubate the tubes at 55°C for 50 mins for the extension step. The 42°C – 60°C temperature range may be the optimaltemperature for extension.

4. The reaction tube from the Step 3 must be incubated at 70°C for 15 minutes for inactivating the Reverse Transcriptase before amplification

Storage Buffer

The enzyme is supplied in a storage buffer consisting of 20 mM Tris-HCl (pH 7.4), 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, 0.01% (v/v) NP-40 and 50% (v/v) glycerol.

Required Materials but Not Provided
  • Vortex or equivalent
  • Microcentrifuge
  • PCR tubes for your instruments
  • Ice water bath
  • Temperature-controlled water bath or heat blocks; the thermal cycler can also be used.
Performance

The stability RT-qPCR data shows the performance of RScript reverse transcriptase is maintained without any significant alteration in performance at 37 °C after a period of 3 weeks (21 days), demonstrating its high resistance to temperature and time. The percentage of RScript reverse transcriptase activity was calculated by dividing values at each reaction temperature.

RScript Reverse Transcriptase

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