|
Product Description
Moloney Murine Leukemia Virus (M-MuLV) Reverse Transcriptase
is a DNA polymerase which utilizes RNA as a substrate. This enzyme can perform cDNA synthesis by
extending off a DNA primer annealed to an RNA template, or can copy a single-stranded DNA template.
M-MuLV Reverse Transcriptase is completely deficient in 3’-5’ proofreading exonuclease function.
Applications
cDNA synthesis from single-stranded RNA or DNA.
Multiple transcripts can be analyzed from a single RT-reaction.
Source of Protein
A recombinant E. coli strain carrying the Moloney
Murine Leukemia Virus Reverse Transcriptase gene.
Components
1. M-MuLV Reverse Transcriptase supplied in 10 mM Tris-HCl, pH 7.6, 150 mM NaCl,
1 mM DTT, 0.1 mM EDTA, 0.1% NP40 alternative, 50% Glycerol.
2. 10X Reaction Buffer containing 500 mM Tris-HCl, pH 8.3, 30 mM MgCl2, 750 mM KCl, 100 mM DTT,
1 mM EDTA.
Storage Conditions
Store at –20°C.
Concentration
200 units/µL
Unit Definition
1 unit is defined as the amount of enzyme required to incorporate
1 nmol of dTTP into acid insoluble material in 10 minutes at 37°C using poly r(A)/oligo (dT) as
a substrate.
Quality Assurance
Purified free of contaminating endonucleases and exonucleases.
In addition, enzyme purity is analyzed by SDS-PAGE, and negligible E.coli genomic DNA is confirmed
by qPCR.
PCR Guidelines
Taq DNA Polymerase is the original and most commonly used PCR enzyme. Taq excels at amplifying shorter (<5 kb) sequences
from low-complexity template sources and produces robust yields with little or no optimization of reaction conditions.
Suggested Reaction Conditions
Incubate at 37°C with 1X Reaction Buffer supplemented with 100 μM dNTPs (not included).
|
