EnzScript (Rev Trans) (10000 U)

A QIAGEN OEM reverse transcriptase engineered as an M-MLV RNase H-minus variant for first-strand cDNA synthesis. Three point mutations in the RNase H domain eliminate detectable RNase H activity and increase the enzyme's thermostability relative to wild-type M-MLV, supporting greater cDNA yield of full-length transcripts. The enzyme generates first-strand cDNA from poly(A) mRNA or total RNA for downstream applications including RT-PCR, cDNA cloning, and library construction for RNA-Seq. The manufacturer cites a recommended input of 50 ng to 2 µg RNA for end-point PCR, suitability for one-step RT-qPCR and long-range RT-PCR, and a working temperature range of 37°C to 50°C, with a stated optimum extension temperature of 42°C.

The protein is expressed in a recombinant E. coli strain carrying the Moloney-Murine Leukemia Virus reverse transcriptase gene. Stated physical purity is >95% by SDS-PAGE with silver staining. Quality-control release criteria report: single-stranded exonuclease <5.0% release at 2,000 U tested, double-stranded exonuclease <1.0% release at 2,000 U tested, double-stranded endonuclease = no conversion at 2,000 U tested, E. coli DNA contamination <10 copies at 2,000 U tested, and no detectable non-specific RNase contamination. The functional RT-PCR assay confirms synthesis of a 9.4 kb cDNA transcript. One unit is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insoluble material in 10 minutes at 37°C using poly r(A) and oligo(dT) as substrate. The reported molecular weight is 75.9 kDa.

The enzyme is supplied in 20 mM Tris-HCl, 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, <0.01% NP-40, and 50% glycerol at pH 7.5 at 25°C, and is provided with a 5X M-MLV Reverse Transcriptase RNase H- Reaction Buffer and 100 mM DTT. Recommended storage is between -25°C and -15°C.