10,000 U of EnzScript Reverse Transcriptase (0.05 mL at 200,000 U/mL) and 5x M-MLV Reverse Transcriptase RNase H Minus Buffer (1 x 1.5 mL) and 100 mM DTT (1 x 1.5 mL)
The EnzScript Reverse Transcriptase is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.
Features
- RNase H-deficient M-MLV with increased thermal stability
- Suitable for cDNA cloning and RT-PCR
- Recommended for library construction for RNA-Seq transcriptome studies
- Compatible with random hexamer, oligo(dT) and gene-specific priming
Product Details
EnzScript Reverse Transcriptase is a Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase with point mutations in the RNase H domain that eliminate detectable RNase H activity. EnzScript can be used to generate first-strand cDNA from poly(A) mRNA or total RNA for use in downstream applications such as RT-PCR, cDNA cloning or library construction for RNA-Seq. The point mutations in the RNase H domain increase the thermostability of the enzyme and support greater cDNA yield of full-length transcripts than wild type M-MLV reverse transcriptase.
One unit of EnzScript Reverse Transcriptase is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into acid insoluble material in 10 minutes at 37°C using poly r(A)/oligo(dT) as a substrate.
EnzScript Reverse Transcriptase is supplied in 20 mM Tris-HCl, 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, <0.01% NP-40 stabilizer and 50% glycerol; pH 7.5 at 25°C.
EnzScript Reverse Transcriptase is supplied with 5X M-MLV Reverse Transcriptase RNase H– Reaction Buffer (cat. no. B7601) and 100mM DTT (cat. no. B9060).
Performance
Polymerase properties
- Optimum extension temperature: 42°C
- Transcript length: Up to 9.4 kb
| Test | Specification |
| Purity | >95% |
| Specific activity | 180,000 U/mg |
| Single-stranded exonuclease | 2000 U <5% released |
| Double-stranded exonuclease | 2000 U <1% released |
| Double-stranded endonuclease | 2000 U = No conversion |
| E. coli DNA contamination | 2000 U <10 copies |
| RNase contamination | 2000 U; No detectable non-specific RNase |
| Functional RT-PCR assay | Synthesis of 9.4 kb cDNA transcript |