M-MLV (Moloney Murine Leukemia Virus) Reverse Transcriptase (RT) is a recombinant DNA polymerase that synthesizes a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. Compared to AMV RT, M-MLV RT lacks DNA endonuclease activity and has a lower RNase H activity.
• Thermostability – Optimal activity at 37°C
• Size of cDNA – MMLV can be used to synthesize first-strand cDNA up to 7 kb
• Applications – Synthesis of first-strand cDNA (1), primer extension (2), sequencing dsDNA (3), cDNA libraries, and RT-PCR (4)
Source: Purified from E. coli expressing the pol gene of M-MLV on a plasmid (5).
Performance and Quality Testing: SDS-PAGE purity; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product.
Unit Definition: One unit of M-MLV RT is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min. at 37°C using poly(A)
• oligo(dT)12-18 as template
• primer (6).
Unit Reaction Conditions: 50 mM Tris-HCl (pH 8.3), 40 mM KCl, 6 mM MgCl2, 1 mM DTT, 0.5 mM [3H]dTTP, 0.1 mM poly(A), 0.1 mM oligo(dT)12-18, 0.1 mg/ml BSA, and enzyme in 50 µl for 10 min. at 37°C (6).
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.