OneScript® Hot cDNA Synthesis Kit
Cat. No.
G594
Unit
100 rxn
Price
$207.00
| Cat. No. | G594 | |||||||||||||||||
| Name | OneScript® Hot cDNA Synthesis Kit | |||||||||||||||||
| Unit | 100 rxn | |||||||||||||||||
| Category | Reverse Transcriptase & RT-PCR | |||||||||||||||||
| Description |
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:
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| Storage Condition |
Store at -20°C. |
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| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G594 |
| Can I use both total RNA and poly(A)+ mRNA for cDNA synthesis? | |
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Yes, both total RNA and poly(A)+ mRNA can be used, though poly(A)+ mRNA typically yields higher quantities and better purity.
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| Can I synthesize cDNA from long RNA transcripts? | |
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Yes, for longer RNA transcripts, extend the 60°C incubation time to up to 30 minutes.
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| What can I use the synthesized cDNA for? | |
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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.
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| How should I store the synthesized cDNA? | |
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Store the synthesized first-strand cDNA at -20°C for long-term use.
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| Why am I getting low cDNA yields? | |
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Low cDNA yields can be caused by poor RNA integrity, contamination, or insufficient RNA input. To improve yield:
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| How much RNA template should I use for cDNA synthesis? | |
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We recommend using 1 ng to 2 μg of RNA per reaction.
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| What volumes of cDNA should I use for downstream PCR? | |
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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.
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What if input RNA samples are expected to have high levels of genomic DNA contamination?
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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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| What is the advantage of OneScript® Hot Reverse Transcriptase compared to OneScript® Plus Reverse Transcriptase? | |
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OneScript® Hot Reverse Transcriptase offers a significant advantage over enzymes that operate at lower temperatures (42-55ºC) due to its exceptional thermostability, allowing it to function effectively at 60-72ºC. This higher temperature improves efficiency by preventing RNA secondary structures and enabling cDNA synthesis from degraded or low-abundance RNA. Additionally, it is more resistant to inhibitors and contaminants, making it ideal for complex or impure RNA samples.
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Do I need an RNase inhibitor when using this product?
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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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| What is the source of this enzyme? | |
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The enzyme is produced recombinantly in E. coli, which has been engineered to express the enzyme gene. While the original gene may come from another organism, all production and purification occur using E. coli under controlled conditions.
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| Can this product be used for next-generation sequencing (NGS) library preparation? | |
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This product can be used for NGS library preparation, but additional processing steps are recommended. NGS workflows require double-stranded DNA, and the presence of RNA:cDNA hybrids may interfere with downstream applications. To ensure compatibility, we suggest treating the reaction with RNase H to remove any residual RNA, followed by PCR amplification to generate the necessary double-stranded DNA for NGS library construction. |
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