M-MLV Reverse Transcriptase (Glycerol-Free)
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 enzyme dilution buffer. The enzyme formulation does not contain glycerol and is compatible for further lyophilization process.
Package & Component :
Source:
Escherichia coli
Purity:
>98% as determined by SDS-PAGE analysis (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.
Storage:
Stored at -20°C. Avoid repeated freeze/thaw cycles.
Shipping Conditions:
Blue ice
| Cat. | Name | Amount |
| C15032-20000U | M-MLV Reverse Transcriptase (Glycerol-Free) (200 U/μL) | 20,000 U |
| C15032-50000U | M-MLV Reverse Transcriptase (Glycerol-Free) (200 U/μL) | 50,000 U |
Source:
Escherichia coli
Purity:
>98% as determined by SDS-PAGE analysis (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.
Storage:
Stored at -20°C. Avoid repeated freeze/thaw cycles.
Shipping Conditions:
Blue ice
The following procedure is a general guideline for RT-qPCR reaction. To maintain an RNase-free environment, always wear disposable gloves, and use laboratory consumables and water of nuclease-free grade during the whole experiment course.
RT-qPCR reaction set-up:
1. Place all required reagents on ice.
*See Usage Notes for additional guidelines on primer/template preparation.
2. Gently mix the reaction thoroughly to achieve uniform distribution and briefly centrifuge.
3. Thermal cycling conditions for standard qPCR.
RT-qPCR reaction set-up:
1. Place all required reagents on ice.
| Component | Amount | Final concentration |
| 2X Probe qPCR Master Mix | 10 μL | 1 X |
| M-MLV Reverse Transcriptase (Glycerol-Free) |
1 μL | 200 U/rxn |
| Forward primer (10 μM) | 0.8 μL | 0.4 μM |
| Reverse primer (10 μM) | 0.8 μL | 0.4 μM |
| Probe (10 μM) | 0.4 μL | 0.2 μM |
| RNA template | X μL | ≦1 μg (total) |
| Nuclease-Free H2O | Y μL | - |
| Total reaction volume | 20 μL | - |
2. Gently mix the reaction thoroughly to achieve uniform distribution and briefly centrifuge.
3. Thermal cycling conditions for standard qPCR.
| Step | Cycles | Temperature | Time |
| Reverse transcription | 1 | 42-50°C | 10–15 min |
| Enzyme activation | 1 | 95°C | 5 min |
| Denaturation | 40-45 | 95°C | 5–15 sec |
| Annealing/Extension | 55-65°C | 30–60 sec |