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What is the recommended concentration for EpiScript RNase H- Reverse Transcriptase in RT reactions?

221, RDXERT300

For 1-step RT-qPCR, we have found 20-50 units per reaction to work best. In 2-step RT-qPCR, we have found 5-20 units per reaction to work best.

What is ‘template-switching’, and does EpiScript RNase H- Reverse Transcriptase have this capability?

221, RDXERT300

Template-switching is the addition of non-template nucleotides to the 3′ end of the transcript, followed by annealing of a primer. This is followed by second strand synthesis by the reverse transcriptase using its innate DNA polymerase activity. EpiScript RNase H- Reverse Transcriptase does have this capability.

What is the difference between EpiScript™ RNase H- Reverse Transcriptase and EpiScriptT RT PLUS?

AS3107, MBTOOL-007, PCRAMP-104

EpiScript RT PLUS contains a blocking mechanism that suppresses reverse transcriptase activity at lower temperatures allowing for room temperature reaction setup.

Can I use EpiScript RNase H- Reverse Transcriptase for monitoring gene expression in mammalian cells?

221, RDXERT300

EpiScript RNase H- Reverse Transcriptase is very useful in the monitoring of gene expression and can be used with both probe hydrolysis and gene intercalation methods of detection.

What conditions do you recommend for set up of RT reactions using EpiScript RNase H- Reverse Transcriptase?

221, RDXERT300

For best results we recommend preparing reactions on ice for both 1-step and 2-step RT-qPCR experiments.

I left the RapiDxFire Thermostable Reverse Transcriptase (RT) out on the bench overnight. Is it still good?

307

Yes, the RapiDxFire RT is stable at room temperature (20 °C) for at least three months. However, we have not tested the stability of the combined reagents (buffer, template, primers) over time at room temperature.

How should I extract or purify my RNA samples if I wish to reverse transcribe them using EpiScript RT PLUS?

AS3107, MBTOOL-007, PCRAMP-104

MasterPure™, sbeadex™ and QuickExtract™ isolation chemistries are all suitable for use with EpiScript RT PLUS.

How should I extract or purify my RNA samples if I wish to reverse transcribe them using Lyo-Ready EpiScript Thermostable RT?

30260-1
Column-based and magnetic bead-based isolation chemistries are both suitable for use with Lyo-Ready EpiScript™ Thermostable Reverse Transcriptase.

Which RT enzyme (MMLV, EpiScript RNase H- Reverse Transcriptase, or EpiScript RT PLUS) should I use for pathogen detection, gene expression, or RNA-Seq library preparation, and why?

AS3107, MBTOOL-007, PCRAMP-104

All three reverse transcriptases; standard MMLV, EpiScript RNase H- Reverse Transcriptase and EpiScript RT PLUS can be used for these applications. EpiScript RNase H- Reverse Transcriptase has the advantage of being active up to 55 °C making it more specific than standard MMLV RT enzymes. EpiScript RT PLUS provides the added benefit of allowing room temperature reaction setup.

Which RT enzyme, MMLV or EpiScript RNase H- Reverse Transcriptase, should I use for pathogen detection, gene expression, RNA-Seq library preparation for the human health care market and why?

221, RDXERT300

Both standard MMLV and EpiScript RNase H- Reverse Transcriptase can be used for these human health care applications. EpiScript RNase H- Reverse Transcriptase has the advantage of being active up to 55 °C making it more specific than standard MMLV RT enzymes.

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