RScript cDNA Synthesis Kit

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

RScript cDNA Synthesis Kit-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-is the latest addition to our reverse transcription product boutique. The RScript cDNA Synthesis Kit contains 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

Primer amounts recommended for efficient cDNA synthesis are 2.5 μM of oligo(dT) (anneal to the 3'-poly(A) + mRNA) or 2.5 ng/μl of random primers (anneal at non-specific sites of RNA templates), or 2 μM of genespecific primers per 20 μl reaction.

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.

*RNA template, primer and 2X Sharp Reaction Mix 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 optimal temperature 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.

• 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.

Reverse transcription of a 330 bp fragment CCD1A RNA using RScript cDNA Synthesis Kit to carry this experiment. The PCR data shows that RScript cDNA Synthesis Kit have a stable and superior performance at higher temperatures.