ThermoScript™ RT-PCR System for First-Strand cDNA Synthesis - FAQs

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3 product FAQs found

How long can I store the cDNA from my reverse transcription step?

You can store your cDNA at 2-6 degrees C for up to 24 hours. For long-term storage, store the cDNA at -15 to -25 degrees C and add EDTA to a final concentration of 1 mM to prevent degradation.

My mRNA has a high-GC content. Which reverse transcriptase do you recommend using?

We would recommend using SuperScript IV Reverse Transcriptase, which offers high thermostability and can be used in reactions at 50-65 degrees C to facilitate the successful transcription of challenging templates (high-GC content or extensive secondary structures).

What is the highest temperature that MMLV, SuperScript II, SuperScript III, or SuperScript IV RTs can be used?

The optimal temperature for for MMLV is 42 degrees C. The optimal temperature for SuperScript II RT is 42 degrees C, and can be used up to 50 degrees C. The optimal temperature for SuperScript III RT is 50 degrees C, and can be used up to 55 degrees C. For some qRT-PCR reactions where gene-specific primers are used, you can do the RT reaction at 60 degrees C. The optimal temperature for SuperScript IV RT is 50 degrees C, but can be used up to 65 degrees C.