货号： 28025013

规格

内容及储存

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.

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).

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.