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Catalog Number | No. of Reactions |
---|---|
28025013 | 200 Reactions |
28025021 | 1,000 Reactions |
M-MLV Reverse Transcriptase 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, Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) lacks DNA endonuclease activity and has a lower RNase H activity.
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)25 as template-primer.
Features of M-MLV Reverse Transcriptase include:
• Excellent thermostability with optimal activity at 37°C
• Can be used to synthesize first-strand cDNA up to 7 kb
• Multiple applications including synthesis of first-strand cDNA, primer extension, sequencing dsDNA, cDNA libraries, and RT-PCR
Source
Purified from E. coli expressing the pol gene of M-MLV on a plasmid
Performance and quality testing
SDS-PAGE purity; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product
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)25, 0.1 mg/mL BSA, and enzyme in 50 μL for 10 min at 37°C
• M-mLV RT, 200 μL (200 U/μL)
• 5X First Strand Buffer, 1 mL
• DTT, 500 μL (100 mM)
Store at –20°C.