M-MLV (H-) Reverse Transcriptase is an RNA-dependent DNA polymerase with reduced RNase H activity. This enzyme can use RNA (when synthesizing cDNA) or single-stranded DNA as a template and initiate the synthesis of a complementary DNA strand from a primer. M-MLV(H-) reverse transcriptase has no 3´ → 5´ exonuclease activity. It can be used to synthesize first strand cDNA more efficient than the wild type M-MuLV。

Storage Solution : 100 U/ul M-MLV(H-) Reverse Transcriptase、20 mM Tris-HCl、100 mM NaCl、1 mM DTT、0.1 mM EDTA、0.01% Nonidet® P-40、50% Glycerol pH 7.5 @ 25°C
5*Reaction Buffer: 250 mM Tris-HCl、375 mM KCl、15 mM MgCl2、50 mM DTT (pH 8.3 at 25°C)

1. Genomic DNA was removed from the extracted cell total RNA by DNase I.
2. After incubating at 37 °C for 30 minutes, add 1µl 0.5 M EDTA (to the final concentration of 5mm) and inactivate at 75°C for 10 minutes.
3. Mix RNA sample, Random Primer and 1 ul M-MLV(H-) Reverse Transcriptase in a sterile RNase-free microfuge tube.
4. Incubate the 20 μl cDNA synthesis reaction was incubated at 25℃ for 5 minutes and then at 42℃ for 1 hour.
5. Inactivate the enzyme at 65°C for 20 minutes. The cDNA product can be directly fed into the qPCR reaction or stored at -20°C.

Please avoid repeated freeze-thaw cycles.