描述

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).

規格

内容及儲存

M-MLV RT is supplied with a vial of 5X first strand buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2 ], and a vial of 100 mM DTT. Store at -20°C. Guaranteed stable for 6 months when properly stored.