The Dongsheng M-MLV Reverse Transcriptase (RT) is a genetically modified M-MLV RT. It differs from the M-MLV RT by its structure and catalytic properties. The enzyme possesses an RNA-dependent and DNA-dependent polymerase activity and a ribonuclease H activity specific to RNA in RNA-DNA hybrids. Dongsheng M-MLV Reverse Transcriptase has significantly lower RNase H activity than Avian Myeloblastosis Virus (AMV) reverse transcriptase.
Source
E.coli cells with a cloned fragment of the pol gene encoding Moloney Murine Leukemia Virus reverse transcriptase.
Applications
Generation of first strand cDNA for use in:
-PCR, see Protocol for First-strand cDNA Synthesis;
-real-time PCR;
-second strand cDNA synthesis;
DNA labeling;
Analysis of RNA by primer extension.
Quality Control
The absence of endodeoxyribonucleases, exodeoxyribonucleases, phosphatases and ribonucleases confirmed by appropriate quality tests.
Functionally tested in first strand cDNA synthesis.
Store Buffer
20m M Tris-HCl (pH 7.5)
200 mM NaCl
0.1 mM EDTA
1 mM DTT
0.01% NP-40
50% glycerol
5xFirst-Strand Buffer
250 mM Tris-HCl (pH 8.3 at 25°C)
375 mM KCl
15 mM MgCl2
50 mM DTT
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