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Thermo Scientific™ Maxima H Minus Double-Stranded cDNA Synthesis Kit
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Thermo Scientific Maxima H Minus Double-Stranded cDNA Synthesis Kit is a complete system for efficient synthesis of double-stranded cDNA from total RNA or mRNA. First- and second-strand cDNA synthesis reactions are performed in the same tube without the need for intermediate organic extraction or ethanol precipitation steps. This convenient single-tube format speeds up the synthesis procedure and maximizes cDNA recovery. The kit contains premixed components to reduce the number of pipetting steps necessary to complete the procedure.
First- and second-strand cDNA synthesis reactions are performed in the same tube without the need for intermediate organic extraction or ethanol precipitation steps. This convenient single-tube format speeds up the synthesis procedure and maximizes cDNA recovery. The kit contains pre-mixed components to reduce the number of pipetting steps necessary to complete the procedure.
Features of the Maxima H Minus Double-Stranded cDNA Synthesis Kit include:
- Efficient synthesis of full-length double-stranded cDNA
- Fast—procedure completed in less than two hours
- Convenient—pre-mixed components
- Complete—includes all primers, controls and residual RNA removal reagents
Applications
- Full-length double-stranded blunt-end cDNA synthesis for cloning
- Double-stranded cDNA library construction
Specifications
Specifications
| Content And Storage | -25°C to -15°C |
| Format | 50 reactions |
| Optimal Reaction Temperature | 50°C to 55°C |
| Product Line | Maxima |
| Product Type | cDNA Synthesis Kit |
| Quantity | 50 Reactions |
| Reverse Transcriptase | Maxima H Minus |
| For Use With (Application) | Cloning |
| Final Product Type | Double Stranded cDNA |
| No. of Reactions | 50 Reactions |
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Frequently Asked Questions (FAQs)
Trace amounts of reagents used in RNA purification protocols may remain in solution and inhibit first-strand synthesis, e.g., SDS, EDTA, guanidine salts, phosphate, pyrophosphate, polyamines, spermidine. To remove trace contaminants, we recommend re-precipitating the RNA with ethanol and washing the pellet with 75% ethanol, or re-purifying the RNA.
RNA purity and integrity are essential for synthesis and quantification of cDNA. Always assess the integrity of RNA prior to cDNA synthesis. Use freshly prepared RNA. Multiple freeze/thaw cycles of the RNA sample and synthesized cDNA is not recommended. Avoid RNase contamination and discard low quality RNA.
It is generally beneficial to minimize RNase H activity when aiming to produce long transcripts for cDNA cloning. RNase H degrades RNA from RNA-DNA duplexes, which can result in truncated cDNA during reverse transcription of long mRNA. We also recommended using RNase H Minus RTs for template-independent addition of C nucleotides. In contrast, reverse transcriptases with intrinsic RNase H activity are often favored in 1-step RT-qPCR applications.
The fidelity of RevertAid and Maxima reverse transcriptases is the same as that of wild-type M-MuLV RT, which is in the range of 1 error per 15,000-27,000 nucleotides synthesized.
All Thermo Scientific reverse transcriptases (RevertAid RT, RevertAid H Minus RT, Maxima RT, and Maxima H Minus RT) possess intrinsic TdT activity, although at varying degrees depending upon the reaction conditions. We recommend specialized SuperScript IV Template Switching RT Master Mix for high efficiency in applications requiring template switching RT.
For Research Use Only. Not for use in diagnostic procedures.
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