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M-MuLV Reverse Transcriptase

By MP Biomedicals

M-MuLV Reverse Transcriptase

High efficiency enzyme from Moloney Murine Leukemia Virus

  • Synthesis of full length copies of cDNA >10 kb
  • Low RNase H activity
  • Recombinant enzyme

M-MuLV Reverse Transcriptase, a recombinant enzyme, is an RNA-dependent DNA polymerase. It is a single polypeptide with an apparent molecular weight of 71,000 Da. It requires Mg2+ for optimal activity. As M-MuLV RT has a lower Rnase H activity than AMV reverse transcriptase it is a better choice for synthesizing full-length copies of large mRNA (in excess of 10 kb), under optimal reaction conditions. It is recommended that 200-300 units of M-MuLV reverse transcriptase are used per µg of mRNA in a reaction at 37°C for 1 hour. This enzyme can synthesize a complementary strand initiating from a single stranded RNA or DNA template. Applications include: synthesizing cDNA for cloning or for hybridization probes, filling in and labeling the 3'-termini of DNA with 5'-protruding ends, DNA sequencing, and amplification of RNA.

References:
Chen, E.Y. and Seeburg, P.H. (1985) DNA 4:165
Heideker, G. and Messing, (1987) J., Methods Enzymol. 154:28
Houts, G.E. et al., (1979) J.Virol.29:517
Roth, M.J., et al. (1985) J. Biol. Chem. 260:9326

Storage: 50 mM Tris HCl, pH 8.3, 0.1M NaCl, 1 mM EDTA , 5 mM DTT, 0.1% Triton X-100 , 50% Glycerol (v/v). Store at -20 °C
Incubation buffer 1x: 50 mM Tris HCl pH 8.3, 75 KCl, 3 mM MgCl2, 10mM DTT .
Unit definition: One unit incorporates one nmol of dTTP into DE-81 adsorbable form in 10 minutes at 37°C using polyA-oligo dT12-18 as a substrate.
Purity: Free of contaminating, non-specific double-stranded and single stranded endonucleases, exonucleases and ribonucleases. Typically greater than 90% pure by SDS-polyacrylamide gel electrophoresis.
Activity: 20-40 units/µL
cDNA synthesis activity: This M-MuLV RT exhibits superior stability during incubation at 37°C. The amount of enzyme required is generally only around 10 units per µg of mRNA.
Concentration: 20-40U/µL

Supplied with 5x incubation buffer and DTT 100mM apart

Storage: -20°C

           
Cat.No. Name Size EA    
17-EMMLV100 M-MuLV Reverse Transcriptase 1000 U $78.24
1/EA
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17-EMMLV103 M-MuLV Reverse Transcriptase 3 x 1000 U $213.63
1/EA
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