Stop-Start RT
Hot start MMLV reverse transcriptase
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Stop-Start reverse transcriptase is an MMLV RT coupled with an reversibly-bound aptamer that inhibits RT activity at room temperature by blocking the nucleic acid binding site of the enzyme, which significantly reduces the formation of non-specific PCR products.
- Compatible with 1-step RT-PCR and 1-step RT-qPCR
For research use only (RUO). Not for use in diagnostic procedures.
Additional information
| Size | 50 reactions, 250 reactions |
|---|---|
| Price | $49, $249 |
| SKU | PQ.0012.Kit.96.RUO |
Overview
Moloney Murine Leukemia Virus (M-MuLV, MMLV) Reverse Transcriptase is an RNA-directed DNA polymerase. This enzyme can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. (1-4). M-MuLV Reverse Transcriptase lacks 3´ → 5´ exonuclease activity.
As shown here, the aptamer has a dose-dependent effect on MMLV RT activity, with non-specific products – and potential false positives amplicons – being eliminated in a one-step RT-PCR.
Kit Contents
50 Reactions
| Contents | Volume (µL) |
| Stop-Start RT | 50 µL |
| 5x Reaction Buffer | 200 µL |
| 25mM MgCl2 | 1,000 µL |
250 Reactions
| Contents | Volume (µL) |
| Stop-Start RT | 250 µL |
| 5x Reaction Buffer | 1,000 µL |
| 25mM MgCl2 | 1,000 µL |
Resources
User guides and protocols
General cDNA Synthesis Protocol
| Reaction Component | Volume (µL) |
| 5x Reaction Buffer | 4 µL |
| Stop-Start RT [5U / µL] | 1 µL |
| MgCl2 [1.5–5.0mM] | variable µL |
| cDNA synthesis primer | variable µL |
| RNase Inhibitor [10-20U – optional] | variable µL |
| Nuclease-free H2O | to final volume of 20 µL |
| Final Volume | 20 µL |