M-MLV Reverse Transcriptase (RNase H-) can synthesize a complementary DNA strand using single-stranded RNA or DNA as a template. This product is a mutant strain cloned through genetic recombination, in which the RNase H activity has been removed while further enhancing thermal stability. The enzyme can perform the synthesis of the first strand of cDNA at 55-60°C and is suitable for reverse transcription of RNA templates with complex secondary structures. It exhibits improved efficiency in reverse transcription and the ability to transcribe long template fragments, making it suitable for low-template and low-copy gene reverse transcription, with the capacity to synthesize cDNA of approximately 10 kb in length.
Components
M-MLV Reverse Transcriptase (RNase H-):
Product Number: MMLV-200 µL
- Size: 50 µL of M-MLV Reverse Transcriptase with 200µL of 5× MMLV RT Buffer
- Number of Reactions: 50 reactions
Product Number: MMLV-800 µL
- Size: 200 µL of M-MLV Reverse Transcriptase with 800µL of 5× MMLV RT Buffer
- Number of Reactions: 200 reactions
Product Number: MMLV-2mL
- Size: 1mL of M-MLV Reverse Transcriptase with 2mL of 5× MMLV RT Buffer
- Number of Reactions: 1000 reactions
- Concentration: 200 units per microliter (200 U/µL)
- Storage Conditions: Store the product at a temperature of -20°C for up to two years.
- Transportation: Transport the product at a temperature of 0°C or lower (≤0°C).
- Incubation: Incubate the product for 10 minutes at a temperature of 37°C.
- Inactivation: Heat the product at 85°C for 5 minutes to inactivate the reverse transcriptase enzyme.
5× MMLV RT Buffer:
- Storage Conditions: Store the product at a temperature of -20°C for up to two years.
- Transportation: Transport the product at a temperature of 0°C or lower (≤0°C).
Compositions:
Enzyme Storage Buffer |
20 mM Tris-HCl (pH 7.5) |
200 mM NaCl |
0.1 mM EDTA |
5 mM DTT |
50% glycerol. |
0.01% NP40 |
5× MMLV RT Buffer |
100 mM Tris-HCl (pH 8.3) |
375 mM NaCl |
15 mM MgCl2 |
50 mM DTT. |
Product Applications: This product is widely applicable for reverse transcription reactions of RNA from animals, plants, and microorganisms.
Definition of Activity: One unit (U) of activity is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insoluble material within 10 minutes at 37°C using Poly(A)·Oligo(dT) as a template-primer.
Quality Control Exonuclease Activity Assay: Incubate 200 U of the enzyme with 1 μg of Hind III-treated λDNA at 37°C for 10 hours. No changes should occur in the DNA electrophoresis pattern.
Endonuclease Activity Assay: Incubate 200 U of the enzyme with 1 μg of pUC19 plasmid DNA at 37°C for 1 hour. No changes should occur in the DNA electrophoresis pattern.
Ribonuclease Activity Assay: Incubate 200 U of the enzyme with 1 μg of 16S, 23S rRNA at 37°C for 1 hour. No changes should occur in the RNA electrophoresis pattern.
Please see manual for more information.
