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| Product Name | >98% Hot-Start MMLV Reverse Transcriptase for 20 µL RT Reactions |
| Catalog No. | IEC-HMM-0218 |
| Description | Alta DiagnoTech hot-start reverse transcriptase is derived from Moloney murine leukemia virus and uses reversible modification for room-temperature activity suppression. |
| Intended Use | For high-temperature reverse transcription, one-step RT-qPCR and RNA premix reagent formulation. |
| Principle / Technology | Reversible modification suppresses enzyme activity at room temperature and permits activity at 55°C without a separate activation step. |
| Sample Type | Total RNA: 10 pg to 5 µg; poly(A) RNA: 10 pg to 500 ng. |
| Reaction Conditions / Protocol | For a 20 µL reaction, combine RNA template, 1 µL oligo(dT)18 at 5 µM or random primer mix at 50 µM, 1 µL dNTP mix at 10 mM each, 1 µL RNase inhibitor at 40 U/µL, 1 µL reverse transcriptase at 200 U/µL, 4 µL 5× reverse-transcription buffer and nuclease-free water to volume. Incubate at 50-55°C for 5-30 minutes, then at 85°C for 2 minutes. |
| Components / Formulation | Reverse transcriptase and 5× reverse-transcription buffer. Enzyme volumes are 0.05 mL, 0.5 mL and 10 mL, supplied with matching buffer volumes of 0.5 mL, 5 mL and 10 × 10 mL. |
| Storage Conditions | Store at -25°C to -15°C. |
| Package Specifications | Enzyme volumes: 0.05 mL, 0.5 mL and 10 mL. |
| Quality Control | No detectable endonuclease, exonuclease, nickase or RNase effect after 200 U enzyme is incubated with the specified nucleic-acid substrates at 37°C for 16 hours; residual E. coli genomic DNA is below 1 copy per 200 U by 16S rDNA-specific qPCR. |
| Purity | >98% by SDS-PAGE |
| Activity / Unit Definition | One unit incorporates 1 nmol dTTP into acid-insoluble material in 10 minutes at 37°C using poly(rA)-oligo(dT) as template-primer. |
| Source / Origin | Moloney murine leukemia virus |
| Shipping Conditions | Ship at 0°C or below. |
| Application Notes / Precautions | Use an RNase-free workspace, tubes and tips, and wear clean gloves and a mask. Store cDNA at -20°C or -80°C or use it immediately; cDNA should not exceed one-tenth of the subsequent PCR volume. |
| Alias | Hot-start MMLV reverse transcriptase |
| Product Type | Hot-start reverse transcriptase |
| Function / Efficacy | Provides thermostable cDNA synthesis with reduced room-temperature background activity. |
| Optimal Temperature | 50-55°C reaction temperature; activity is available at 55°C. |
For research use only, not for clinical use.
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