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| Product Name | Ultra Reverse Transcriptase (200 U/μL) |
| Catalog No. | IVDRM-0105 |
| Description | Ultra Reverse Transcriptase (200 U/μL) is a novel enzyme obtained by genetic engineering of an RNase H-deficient M-MLV reverse transcriptase. Lacking RNase H activity, it offers markedly improved full-length cDNA synthesis and can amplify cDNA up to 20 kb. Its significantly enhanced thermostability permits reactions at temperatures up to 65°C, enabling efficient reverse transcription of RNA templates with complex secondary structures. Compared with the parental M-MLV (H-) enzyme, it provides higher processivity and synthesis rates together with increased tolerance to inhibitors, effectively overcoming low reverse transcription efficiency caused by inhibitors present in samples. |
| Intended Use | Full-length cDNA library construction; endpoint PCR; RT-PCR and real-time quantitative RT-PCR. |
| Principle / Technology | Genetically engineered M-MLV (H-) reverse transcriptase lacking RNase H activity; enhanced thermostability (up to 65°C), processivity, and inhibitor tolerance; possesses terminal transferase (TdT) activity that adds nucleotides to the 3' end of synthesized DNA in a template-independent manner. |
| Sample Type | Total RNA and other RNA templates, including samples containing reverse transcription inhibitors. |
| Performance Range / Specifications | Ultra-high sensitivity: efficient reverse transcription of as little as 1 pg total RNA. Fast reverse transcription: cDNA synthesis completed in 5 min. Strong processivity: no RNase H activity; maximum synthesis length up to 20 kb. High thermostability: reaction temperatures up to 65°C, improving compatibility with templates of complex structure. TdT activity: non-template addition of nucleotides to the 3' end of synthesized DNA. |
| Sensitivity / LOD | Efficient reverse transcription of 1 pg total RNA. |
| Reaction Conditions / Protocol | Reverse transcription can be performed at up to 65°C; cDNA synthesis completed within 5 min (e.g., 55°C for 5 min as validated in RT-qPCR workflows). |
| Components / Formulation | Supplied as a standalone enzyme at 200 U/μL. |
| Storage Conditions | Store at -20°C. |
| Shelf Life | 1 year when stored at -20°C. |
| Package Specifications | 2000 U; 10000 U; 200000 U |
| Quality Control | No nicking enzyme or exonuclease contamination: 200 U incubated with 0.5 μg of various substrate DNAs at 37°C for 4 h shows no change in DNA banding by agarose gel electrophoresis. No RNase contamination: 200 U incubated with 0.5 μg substrate RNA at 37°C for 4 h shows no change in RNA banding. RNA-seq library yield exceeds that of a leading international brand, with comparable sequencing quality and lower host DNA residue. |
| Purity | Free of detectable nicking enzyme, exonuclease, and RNase activity. |
| Concentration | 200 U/μL |
| Source / Origin | Genetically engineered from M-MLV (H-) reverse transcriptase. |
| Shipping Conditions | Shipped on dry ice. |
| Expiration Date / Stability | Stable for 1 year at -20°C. |
| Recommended Buffer System | Optimized reverse transcription buffer for RT-PCR/qPCR applications. |
| Application Notes / Precautions | Elevated reaction temperatures (up to 65°C) are recommended for RNA templates with complex secondary structures. The enzyme's enhanced inhibitor tolerance makes it suitable for crude or inhibitor-containing samples. |
| Batch-to-Batch Consistency | Excellent lot-to-lot stability: three independent lots tested with 5, 50, and 500 ng of 293T RNA (reverse transcription at 55°C for 5 min followed by qPCR) showed ΔCt values < 0.5 at every input level. |
For research use only, not for clinical use.
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