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RT-PCR

Promega reverse transcription-PCR (RT-PCR) products include standalone reverse transcriptases (AMV, M-MLV and GoScript) and a range of complete kits containing all the required enzymes, buffers and reagents for successful RT-PCR. 

GoScript™ Reverse Transcriptase includes M-MLV reverse transcriptase and a proprietary buffer designed for robust, reliable cDNA synthesis from a full range of rare and abundant transcripts.  GoScript is available in convenient mixes with either Oligo(dT) primers or random primers, as part of a complete kit, and as a stand-alone enzyme.

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RT-PCR Basics

Reverse transcriptases (RTs) are RNA-directed DNA polymerases that synthesize a DNA copy (cDNA) from a target RNA. To apply PCR to the study of RNA, the RNA sample must first be reverse transcribed to cDNA to provide the necessary DNA template for subsequent PCR.

In RT-PCR, after an initial reverse transcription step to produce the cDNA template, PCR is performed to amplify the target sequence. In one-step RT-PCR, reverse transcription and PCR are performed in a single reaction mix. In two-step RT-PCR the reverse transcription reaction is performed separately, and the cDNA produced is added to a subsequent PCR. 

AMV Reverse Transcriptase is recommended for one-step and two-step RT-PCR, RT-qPCR, reverse transcription of RNAs <5kb and primer extension, particularly if the template RNA has strong secondary structure.

M-MLV Reverse Transcriptase RNase H– point mutant is the enzyme of choice for reverse transcribing long RNAs for cDNA library construction, cDNA probe generation and primer extension.

GoScript™ Reverse Transcriptase is an M-MLV formulation optimized for use in one-step and two-step RT-qPCR and reverse transcription of rare and abundant targets. GoScript performs well in the presence of many common amplification inhibitors.