| Field | Specification |
|---|---|
| Mfr No | |
| Form | Kit |
| Product Type | |
| Storage |
OneScript® Plus cDNA Synthesis Kit contains a Moloney-Murine Leukemia Virus Reverse Transcriptase with genetic modifications to abolish RNase H activity to achieve thermal stability. This special mutant enzyme offers higher cDNA yields, longer cDNA up to 12 kb, and is able to perform under high temperatures (50°C - 55°C), facilitating the elimination of secondary structures associated with GC-rich RNA templates. OneScript® Plus is formulated with abm’s RNaseOFF Ribonuclease Inhibitor, offering improved resistance to oxidation compared to the high oxidation-sensitive human RNase inhibitors. RNaseOFF is stable even under very low concentrations of DTT (< 1 mM), making it the best choice for ultimate RNA protection.
Product Features
- Maximal flexibility in priming – oligo(dT), random primers, or gene-specific primers
- Robust cDNA synthesis from any RNA template
- High reproducibility and excellent yield
- Employs abm's OneScript® Plus Reverse Transcriptase (Cat. No. G237)
- Included in abm's PCR Buffet Program
| Product Component | Quantity |
|---|---|
| 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 |
| OneScript® Plus Reverse Transcriptase | 100 rxn (100 µl) |
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ISO 13485:2016 MDSAP Certified |
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This product is equivalent to RScript Plus cDNA Synthesis Kit (Cat. No. G236EU). |
| Component | Quantity |
|---|---|
| 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 |
| OneScript® Plus Reverse Transcriptase | 100 rxn (100 µl) |
| 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.
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).
All of abm's Reverse Transcriptases include RNaseOFF Ribonuclease Inhibitor, which protects RNA from degradation and is resistant to oxidation, even under low DTT concentrations. Therefore, there’s no need to add an external RNase inhibitor, as RNaseOFF provides optimal RNA protection during reverse transcription.
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