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Description
M-MLV Reverse TranscriptaseProduct Specification Synonyms MMLV RTM MLV Reverse Transcriptase Expression System E. coli Molecular Weight 76 kDa (Reducing) Purity 95% by SDS PAGE Conjugation Unconjugated Tag His Tag Storage Buffer 50 mM Tris HCl150 mM NaCl1 mM DTT0. 1 mM EDTA50% Glycerol0. 1% NP 40pH 7. 6 @ 25C Stability & Storage Store at 25 ~ 15 for 2 years Reference [1] Blain, S. W. , and S. P. Goff . "Differential Effects of Moloney Murine Leukemia Virus Reverse Transcriptase
Product Specification
| Synonyms | MMLV RT，M-MLV Reverse Transcriptase |
| Expression System | E.coli |
| Molecular Weight | 76 kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Conjugation | Unconjugated |
| Tag | His Tag |
| Storage Buffer | 50 mM Tris-HCl、150 mM NaCl、1 mM DTT、0.1 mM EDTA、50% Glycerol、0.1% NP-40、pH 7.6 @ 25°C |
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference | [1] Blain, S. W. , and S. P. Goff . "Differential Effects of Moloney Murine Leukemia Virus Reverse Transcriptase Mutations on RNase H Activity in Mg and Mn." Journal of Biological Chemistry 271.3(1996):1448-54. |
Background
Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) is an RNA-dependent DNA polymerase. The enzyme can use RNA (when synthesizing cDNA) or single-stranded DNA as a template and initiate the synthesis of a complementary DNA strand from a primer. M-MLV reverse transcriptase has no 3´ → 5´ exonuclease activity.
Components
Storage Solution : 200 U/ul M-MLV Reverse Transcriptase、50 mM Tris-HCl、150 mM NaCl、1 mM DTT、0.1 mM EDTA、50% Glycerol、0.1% NP-40、pH 7.6 @ 25°C
10*Reaction Buffer: 500 mM Tris-HCl、750 mM KCl、30 mM MgCl2、100 mM DTT(pH 8.3 @ 25°C)
Protocol
1. Genomic DNA was removed from the extracted cell total RNA by DNase I.
2. After incubating at 37 °C for 30 minutes, add 1µl 0.5 M EDTA (to the final concentration of 5mm) and inactivate at 75°C for 10 minutes.
3. Mix RNA sample, Random Primer and 1 ul M-MLV Reverse Transcriptase in a sterile RNase-free microfuge tube.
4. Incubate the 20 μl cDNA synthesis reaction was incubated at 25℃ for 5 minutes and then at 42℃ for 1 hour.
5. Inactivate the enzyme at 65°C for 20 minutes. The cDNA product can be directly fed into the qPCR reaction or stored at -20°C.
Guidelines
Please avoid repeated freeze-thaw cycles.
Unit Definition
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