M-MLV RTase (M-MLV Reverse Transcriptase)
Moloney Murine Leukemia Virus (M-MLV) Reverse Transcriptase is an RNA-dependent DNA polymerase that synthesizes the first strand of complementary cDNA from a single-stranded RNA template with hybridized primer. This kit features high activity formulation of M-MLV RT and 5X reverse transcription buffer which are capable of full-length cDNA synthesis and high cDNA yields.
Package & Component :
Source:
Escherichia coli
Purity:
>98% as determined by SDS-PAGE (purified by Ni-NTA chromatography).
Unit Definition:
One unit is defined as the amount of the enzyme incorporates 1 nmol of dTTP into acid-insoluble product in 10 minutes at 37°C.
Reaction Condition:
1X M-MLV Reverse Transcriptase Buffer, supplemented with dNTP mix, template and primer, Incubate at 37°C for synthesis of first strand cDNA.
5X M-MLV Reverse Transcriptase Buffer: 250 mM Tris-HCl (pH 8.3), 15 mM MgCl2, 375 mM KCl, and 50 mM DTT.
Storage Buffer:
M-MLV Reverse Transcriptase is supplied in 20 mM Tris-HCl (pH 7.5), 200 mM NaCl, 0.25 mM EDTA, 0.01% NP-40 (v/v), 2.5 mM DTT, 50% glycerol (v/v).
Storage:
Stored at -20°C. Avoid repeated freeze/thaw cycles.
Notes:
After the reaction is complete, M-MLV RTase can be inactivated by incubation at 65°C for 20 minutes.
Shipping Conditions:
Blue ice
| Cat. | Name | Amount |
| C15021-20000U | M-MLV Reverse Transcriptase (200 U/μL) | 20,000 U |
| 5X M-MLV Reverse Transcriptase Reaction Buffer | 1 mL | |
| C15021-50000U | M-MLV Reverse Transcriptase (200 U/μL) | 50,000 U |
| 5X M-MLV Reverse Transcriptase Reaction Buffer | 1 mL |
Source:
Escherichia coli
Purity:
>98% as determined by SDS-PAGE (purified by Ni-NTA chromatography).
Unit Definition:
One unit is defined as the amount of the enzyme incorporates 1 nmol of dTTP into acid-insoluble product in 10 minutes at 37°C.
Reaction Condition:
1X M-MLV Reverse Transcriptase Buffer, supplemented with dNTP mix, template and primer, Incubate at 37°C for synthesis of first strand cDNA.
5X M-MLV Reverse Transcriptase Buffer: 250 mM Tris-HCl (pH 8.3), 15 mM MgCl2, 375 mM KCl, and 50 mM DTT.
Storage Buffer:
M-MLV Reverse Transcriptase is supplied in 20 mM Tris-HCl (pH 7.5), 200 mM NaCl, 0.25 mM EDTA, 0.01% NP-40 (v/v), 2.5 mM DTT, 50% glycerol (v/v).
Storage:
Stored at -20°C. Avoid repeated freeze/thaw cycles.
Notes:
After the reaction is complete, M-MLV RTase can be inactivated by incubation at 65°C for 20 minutes.
Shipping Conditions:
Blue ice
First strand cDNA synthesis:
1. Place all required reagents to a nuclease-free microcentrifuge tube and following the order suggested below.
2. Heat the tube to 65°C for 10 minutes to denature the secondary structure within RNA template. Immediately cool the tube on ice for 1 minute and centrifuge briefly in microcentrifuge. Add the following components to the annealed primer/RNA template, prepare on ice.
3. Incubate at 37°C for 1 hour. The extension temperature may be adjusted from 37°C to 42°C.
4. Inactivate the reaction at 65°C for 20 minutes. The cDNA products should be store at -20°C.
5. Reaction preparations may be scaled up or down proportionately.
1. Place all required reagents to a nuclease-free microcentrifuge tube and following the order suggested below.
| Component | Amount | Final concentration |
| Oligo (dT)12-18 (50 μM) or random primer mix (60 μM) | 1 μL | - |
| Total RNA template | X μL | 1 μg |
| Nuclease-Free H2O | Y μL | - |
| Total reaction volume | 13.5 μL | - |
| Component | Amount | Final concentration |
| 5X Reverse Transcriptase Reaction Buffer | 4 μL | 1X |
| 10 mM dNTPs (each) | 1 μL | 0.5 mM each |
| RNase Inhibitor | X μL | 20 U/rxn |
| M-MLV RTase | 1 μL | 200 U/rxn |
| Nuclease-Free H2O | Y μL | - |
| Total reaction volume | 20 μL | - |
4. Inactivate the reaction at 65°C for 20 minutes. The cDNA products should be store at -20°C.
5. Reaction preparations may be scaled up or down proportionately.
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