Reverse Transcription

Reverse transcription is the process of transcribing RNA molecules into complementary DNA. This cDNA synthesis reaction is mediated by enzymes called reverse transcriptases, or RTs. Moloney murine leukemia virus (M-MLV) reverse transcriptase is one of the RTs used in molecular biology workflows. However, novel engineered RTs are often preferred for enhanced efficiency, improved thermostability, and higher cDNA yields.

Reverse transcription reactions involve multiple steps to synthesize cDNA. Reverse transcriptases exhibit several distinct biochemical activities, predominantly RNA-dependent DNA polymerase activity and RNase H activity. Firstly, reverse transcriptase binds to an RNA template in the presence of an annealed primer. Next, the RNA-dependent DNA polymerase activity of the enzyme synthesizes cDNA by incorporating dNTPs from the reaction mix. This generates a cDNA:RNA hybrid and is referred to as first-strand cDNA synthesis. If reverse transcriptase exhibits RNase H activity (as in wild-type M-MLV RT), the RNA of the cDNA:RNA hybrid is cleaved during first-strand synthesis. In engineered enzymes, RNase H activity is often reduced to prevent premature RNA degradation.

Choose from a variety of reverse transcriptase stand-alone enzymes, master mixes, cDNA synthesis kits, primers, and dNTP solutions to suit your application. Invitrogen SuperScript IV products aid in exceptional cDNA synthesis from any type of template RNA.