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

查看更多产品信息 ThermoScript™ RT-PCR System for First-Strand cDNA Synthesis - FAQs (11146016)

4 个常见问题解答

我的mRNA之中的GC含量较高。您建议采用哪种反转录酶？

建议您使用我们的ThermoScript 反转录酶，在如今市售的反转录酶之中，这种酶具有最高的热稳定性，可以处理您的较棘手模板（高GC含量或过多二级结构）。

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 our ThermoScript Reverse Transcriptase, which provides the highest thermostability of any RT on the market to address your challenging template (high-GC content or extensive secondary structure).

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

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 II RT is 42 degrees C, and can be used up to 50 degrees C. Optimal temperature for MMLV is 42 degrees C. ThermoScript RT shows optimal activity at 60 degrees C, and can be used at temperatures as high as 70 degrees C (for amplicons expected to be 1 kb or less). For PCR products expected to be greater than 1 kb, a maximum first strand synthesis temperature of 60-65 degrees C is suggested. Be sure your first-strand primer anneals at the high temperature, especially when gene-specific primers are used for high-temperature stable reverse transcriptases. We recommend oligo (dT)20 for cDNA synthesis when using an oligo (dT) primer for first-strand synthesis with these enzymes.