M-MuLV Reverse Transcriptase (RNAse H minus)
DATA SHEET
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M-MuLV Reverse transcriptase (RNase H minus)
Cat.# 7P8
Description:
M-MuLV RT, RNase minus (recombinant modified form of the Reverse Transcriptase from the Moloney Murine Leukemia Virus with inactivated RNase H activity) is an RNA-dependent DNA polymerase. The enzyme is a product of a pol gene of M-MuLV and consists of a single subunit with a molecular weight of 69 kDa. M-MuLV RT has no intrinsic RNase H activity. This enzyme does not demonstrate any degradation of the RNA molecule during first strand cDNA synthesis and gives substantially greater yields of first strand cDNA, than obtained with any other reverse transcriptase.
Storage and dilution buffer:
50 mM Tris-HCl, pH 8.0, 0.1 mM EDTA, 1 mM DTT, 100 mM NaCl, 0.5 % Triton X-100, 50 % glycerol.
Unit definition:
One unit of activity is the amount of enzyme required to incorporate 1 nmoles of deoxynucleotide into acid-precipitable material in 10 minutes at 37 °C using poly(rA)-oligo(dT)50 as a template-primer.
Reaction buffer 5x: 250 mM Tris-HCl, pH 8.0, 375 mM KCl, 15 mM MgCl2.
To 1x reaction buffer DTT should be added to a final concentration of 10 mM.
Applications:
First strand DNA synthesis for cloning and hybridization, filling-in and labelling the 3’-terminy of DNA with 5’-protruding ends, DNA sequencing, and RT-PCR.
Quality control:
Activity, SDS-PAGE purity, absence of dsDNases, Rnases, and endonucleases/nickases.
Concentration:
100-200 units/µl
Storage:
-20 °C
Additional product information:
Enzymes and Reagents for molecular biology TechNotes
Other information:
Some applications in which these products may be used are covered by patents issued and applicable in certain countries. Because purchase of these products does not include a license to perform any patented application, users of these products may be required to obtain a patent license, depending on the particular application and country in which the product is used.
