Lyo-Compatible MMLV-RT
Lyo-Compatible MMLV-RT is a high concentration reverse transcriptase and dilution buffer. Lyo-Compatible MMLV-RT is added to a glycerol free qPCR mix to create a Lyo-ready 1-step RT-qPCR mix for subsequent lyophilization.
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Meridian's Lyo-Compatible MMLV-RT
- Highly sensitive cDNA synthesis, even low concentrations of template RNA
- Thermostability up to 45°C, for more challenging transcripts
- High-quality, first-strand cDNA synthesis ideal for most one-step RT-PCR and RT-qPCR applications
- Ideal for creating ambient-temperature stable assays.
Lyo-Compatible MMLV-RT, MDX042
Glycerol-free high concentration reverse transcriptase and dilution buffer. Can be used to create 1-step RT-qPCR mixes for subsequent lyophilization.
Documents & Resources
List of Lot-Specific Enzyme Concentrations
Reverse transcriptase activity:
356.0 U/µL Lot Numbers: EM032-B069810 , EM033-B075830, EM032-B079280, EN055-B075840, EM032-B070090, EM033-B078120, EM033-B079610, EN055-B077220, EM032-B078560 , EM033-B078550, EN054-B069830 , EM033-B072620, EM116-B079090, EN055-B071120
Reverse transcriptase activity:
820.6 U/µL Lot Numbers: EM116-B079670, EM033-B079680, EM032-B079780, EM033-B079960, EN055-B081110, EN055-B081180, EN055-B081460
Reverse transcriptase activity:
676.6 U/µL Lot Numbers: EN055-B095790, EN055-B102520
Reverse transcriptase activity:
591 U/µL Lot Numbers: EN055-B099440, EN054-B085190, EN055-B092750, EN054-B088060, EN054-B088340, EN054-B088440, EN055-B092210, EN055-B095500, EN055-B096450, EN055-B097970, EN055-B097120, EN055-B097760
Reverse transcriptase activity:
501 U/µL Lot Numbers: EN055-B103720, EN055-B103930, EN054-B105000, EN055-B107580, EN055-B118430
966.0 U/µL Lot Numbers: EN055-B127580, EN054-B129970, EN054-B131550
Description
Lyo-Compatible MMLV-RT is a high-concentration, Moloney Murine Leukemia Virus (MMLV) Reverse Transcriptase (RT) with reduced RNase H activity, which exhibits high sensitivity and efficiency for cDNA synthesis and is suitable for incorporation into wet or lyophilized multiplex RT-PCR assays, allowing high sensitivity detection of low copy number RNA targets.
Specifications
| Description | High concentration reverse transcriptase and dilution buffer. Can be used to create 1-step RT-qPCR mixes for subsequent lyophilization. |
| Concentration | > 165 U/µL (see List of Lot-Specific Enzyme Concentrations) |
| Appearance | Clear, colorless solution |
| Application | cDNA synthesis, RT-PCR, PCR, two-step RT-qPCR, one-step RT-qPCR |
| Sample type | RNA |
| Presentation | 2 vials |
| Storage | -20 °C |
| Mix stability | See outer label |
| DNA contamination | None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets. |
| DNase/RNase Contamination | No detectable degradation |
Catalogs & Brochures
Related Products
FAQs: Lyo-compatible MMLV-RT
I want to use viral RNA with very high secondary structure for cDNA synthesis, can I use Lyo-compatible MMLV-RT?
For RNA with very high secondary structure, we recommend using 55C MMLV-RT (MDX117), this is a glycerol-free reverse transcriptase can be used at temperatures up to 60°C which improves the cDNA yield from difficult RNA targets that require higher temperature to denature strong secondary structures.
Can I improve the yield when using Lyo-compatible MMLV-RT?
Avoiding ribonuclease contamination is of primary concern since high quality template RNA is required. You can add RNase Inhibitor (MDX056).
What buffer do you recommend using with Lyo-Compatible MMLV-RT?
The Lyo-Compatible MMLV-RT is supplied with an enzyme dilution buffer, but if you are going to make cDNA, you will also require a buffer that contains 0.2 mM each dATP, dCTP, dGTP & dTTP, 10 mM DTT (dithiothreitol), 25 mM KCl, 3.5 mM MgCl2 and 50 mM Tris–HCl (7.5). However, if you want to do RT-qPCR, we recommend Lyo-Ready 1-Step RT-qPCR Buffer (MDX052).
How is Lyo-Compatible MMLV-RT inactivated?
The reverse transcriptase can be inactivated by adding a chelating agent such as EDTA or heating to 70°C or higher for 10 min.
Does Lyo-Compatible MMLV-RT need a primer?
Yes, to initiate cDNA synthesis, the Lyo-Compatible MMVL-RT reverse transcriptase requires a primer to bind to its complementary sequences on the RNA template and serve as a starting point for synthesis of a new strand. For general cDNA synthesis, random hexamer primers and oligo(dT) primers can be used either separately or mixed. For RT-PCR and RT-qPCR gene-specific primers should be used.
What is the error rate of Lyo-Compatible MMLV-RT?
Unlike eukaryotic replicative polymerases, reverse transcriptase lacks exonuclease activity and so is error prone. MMLV-based reverse transcriptase has been reported to have an error rate in the range of 1.1 × 10−4 to 4.8 × 10−4.
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