| Field | Specification |
|---|---|
| Mfr No | |
| Form | Kit |
| Product Type | |
| Storage |
OneScript® Hot cDNA Synthesis Kit is the first kit in the world that allows for efficient and robust cDNA synthesis at 60ºC. All other cDNA synthesis kits on the market are optimized for the reaction to be performed at 42-55ºC, which is sufficient for most RNA template, but fails on more difficult templates. abm is the only company in the world to have a reverse transcriptase engineered to offer superior cDNA synthesis performance with even the most challenging RNA samples due to its incredible thermostability at 60-72ºC.
The enzyme is a mutational derivative of Moloney-Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) that efficiently reverse transcribes low-abundance or partially degraded RNA and exhibits enhanced resistance to common inhibitors such as reagents used during RNA extraction and contaminants from biological samples. It features genetic modifications that abolish RNase H activity to achieve thermal stability, allowing the RNA template to remain intact during first-strand synthesis for higher cDNA yield and fidelity. Its high processivity and sensitivity enable rapid synthesis of full-length cDNA fragments in a fraction of the time required by leading competitors. OneScript® Hot is formulated with abm’s RNaseOFF Ribonuclease Inhibitor, which provides improved resistance to oxidation compared to traditional, oxidation-sensitive human RNase inhibitors. RNaseOFF remains stable even under very low concentrations of DTT (<1 mM), ensuring superior RNA protection and reliability in demanding experimental conditions.
Product Features:
- Extremely thermostable cDNA synthesis at 60-72ºC
- Transcribes degraded samples and resists inhibitors/contaminants
- High cDNA yields from difficult or ultra-low RNA samples
- Included in abm's PCR Buffet Program
| Product Component | Quantity |
|---|---|
| OneScript® Hot Reverse Transcriptase | 100 rxn (100 µl) |
| 5X RT Buffer | 400 µl |
| Oligo(dT) (10 μM) | 100 µl |
| Random Primers (10 μM) | 100 µl |
| dNTP (10 mM) | 100 µl |
| Nuclease-Free H2O | 2 x 1.0 ml |
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ISO 13485:2016 MDSAP Certified |
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This product is equivalent to RScript Hot CDNA Synthesis Kit (Cat. No. G594EU). |
| Component | Quantity |
|---|---|
| OneScript® Hot Reverse Transcriptase | 100 rxn (100 µl) |
| 5X RT Buffer | 400 µl |
| Oligo(dT) (10 μM) | 100 µl |
| Random Primers (10 μM) | 100 µl |
| dNTP (10 mM) | 100 µl |
| Nuclease-Free H2O | 2 x 1.0 ml |
| Specification | Value |
|---|---|
| Enzyme Type | cDNA Synthesis Kit |
| Format | Kit |
| RT Origin | M-MLV |
| Storage Conditions | Store at -20°C. |
Yes, both total RNA and poly(A)+ mRNA can be used, though poly(A)+ mRNA typically yields higher quantities and better purity.
Yes, for longer RNA transcripts, extend the 60°C incubation time to up to 30 minutes.
The high-quality cDNA can be used in a variety of downstream applications, including gene expression analysis, cloning, and PCR-based assays. BlasTaq™ 2X qPCR Master Mix (Cat. No. G891) is well suited to downstream qPCR applications.
Store the synthesized first-strand cDNA at -20°C for long-term use.
Low cDNA yields can be caused by poor RNA integrity, contamination, or insufficient RNA input. To improve yield: Check RNA integrity with gel electrophoresis or a BioAnalyzer, aiming for a minimum A260/A280 ratio of 1.7 or a RIN above 8. Clean the RNA using ethanol precipitation or lithium chloride to remove contaminants. Purify the RNA with extraction kits or phenol/chloroform to remove proteins. Increase RNA input, especially for low-abundance samples.
We recommend using 1 ng to 2 μg of RNA per reaction.
Typically, use 1 μl of cDNA in a 25 μl PCR reaction. You can add up to 20% of the PCR volume (e.g., 5 μl in a 25 μl PCR), depending on your target and primers.
If input RNA samples are expected to have high levels of genomic DNA contamination, we recommend using abm’s All-In-One 5X RT MasterMix with gDNA Removal (Cat. No. G592).
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