Certificate of Analysis and Data Sheet

Source:
Recombinant E. coli strain

Catalog No.
ENZ-310

Description:

MMLV (Molony Murine Leukemia Virus) Reverse Transcriptase is a DNA polymerase that synthesizes a complementary DNA strands from single-stranded RNA, DNA, or an RNA-DNA hybrid as a template. This recombinant enzyme was purified from E.coli. which carried a modified MMLV-RT gene. Compared to AMV Reverse Transcriptase, this enzyme has a much weaker 5' - 3' ribonuclease H activity, which allows the syntesis of longer cDNAs (>7kb).

Applications:

1. 1st strand cDNA synthesis. 2. Primer extension. 3. RT-PCR.

Physical Appearance:

Sterile Filtered clear solution.

Unit Definition:

One unit is defined as the amount of enzyme required to catalyze the incorporation of 1nmol of deoxyribonucleotide into acid-insoluble forms in 10 minutes at 37^oC, using poly(A)-oligo(dT)12-18 as the template-primer.

Standard cDNA Synthesis Conditions:

50mM Tris-HCl (pH8.3), 75mM KCl, 3mM MgCl2, 10mM DTT, 1.0mM each dATP, dGTP, dCTP, and dTTP, 0.2 ug radom hexamer,1-5ug RNA, 200units M-MLV RT. The reaction volume was 20痞 and the incubation was 45 min at 42^oC.

Concentration:

200 units/痞.

5X Reaaction Buffer:

250mM Tris-HCl (pH8.3), 375mM KCl, 15mM MgCl2, and 50mM DTT.

Formulation:

50mM Tris-HCl, 0.1M NaCl, 0.1% Triton X-100, 2mM DTT, 0.1mM EDTA and 50% glycerol.

Stability:

Stable for 5 days at 10^oC, for longer period of time store at -20^oC.
Please avoid freeze-thaw cycles.

References:

1. Sambrook, J., et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
2. Gerald, G. F. and D'Alessio, J. M. (1993) In Methods in Molecular Biology, Vol. 16, Humana Press, NJ.

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

Latest Publications:

1. Study of MMLV RT- binding with DNA using surface plasmon resonance biosensor.
Acta Biochim Biophys Sin (Shanghai) 2005 Sep;37(9):634-42
2. Incorporation of (alpha-P-borano)-2',3'-dideoxycytidine 5'-triphosphate into DNA by drug-resistant MMLV reverse transcriptase and Taq DNA polymerase.
Nucleosides Nucleotides Nucleic Acids 2003 May-Aug;22(5-8):1651-5
3. Sensitivity of tyrosinase mRNA detection by RT-PCR: rTth DNA polymerase vs. MMLV-RT and AmpliTaq polymerase.
Biotechniques 1996 Apr;20(4):592-600