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| Product Name | AdvanceFast 1st Strand cDNA Synthesis Kit |
| Catalog No. | RESK-0019 |
| Description | AdvanceFast 1st Strand cDNA Synthesis Kit is a rapid reverse transcription kit designed for both PCR amplification and RT-qPCR. Developed from our proven third-generation cDNA synthesis platform, it delivers guaranteed detection rates, specificity, and yields while completing the entire reverse transcription workflow in as little as under 6 minutes—dramatically shortening experimental time. The kit includes a gDNA Digester Mix that removes residual genomic DNA contamination from RNA templates, ensuring more reliable downstream results. Two cDNA synthesis primers—Random Primers N6 and Oligo(dT)18—are provided, and users may choose Random Primers N6, Oligo(dT)18, or gene-specific primers as the reverse transcription primer according to their needs. The resulting single-stranded cDNA can be used directly in downstream PCR or qPCR. A blue tracking dye in the SuperMix visually indicates pipetting progress without affecting reverse transcription efficiency. |
| Intended Use | Rapid first-strand cDNA synthesis from RNA templates for downstream PCR amplification and RT-qPCR analysis, with optional removal of residual genomic DNA contamination. |
| Principle / Technology | The kit uses an engineered fast reverse transcriptase that completes cDNA synthesis in 5 minutes. A separate 5× gDNA Digester Mix enzymatically removes residual genomic DNA from RNA templates before reverse transcription. Flexible priming options—Random Primers N6, Oligo(dT)18, or user-supplied gene-specific primers—support diverse experimental designs. The 4× SuperMix contains a blue tracking dye that marks pipetting progress; testing confirms the dye does not affect reverse transcription efficiency, qPCR Ct values, melt-curve peaks, or Tm values. The enzyme is free of RNase H activity. |
| Detection Method | Downstream fluorescent quantitative PCR (SYBR Green or probe-based) or conventional PCR |
| Sample Type | Total RNA (10 pg–1 μg), e.g., 293T cell total RNA |
| Performance Range / Specifications | Reverse transcription in only 5 minutes; total workflow time as fast as under 6 minutes. Successful reverse transcription across 10 pg–1 μg RNA input. Reverse transcription efficiency comparable or superior to leading competing products, with lower or equivalent Ct values. Synthesizes cDNA fragments up to 14 kb (extend reverse transcription to 20–30 minutes for fragments above 10 kb). Kit stable for 14 days at 4°C; cDNA product stable for 7 days at 37°C and withstands 50 freeze-thaw cycles. Ct value differences of ≤0.2 between 5-minute and longer reverse transcription protocols. |
| Sensitivity / LOD | Reliably reverse transcribes as little as 10 pg of RNA (verified over the 10 pg–1 μg input range with consistent quantitative results). |
| Specificity | The included 5× gDNA Digester Mix removes residual genomic DNA contamination from RNA templates, ensuring reliable downstream quantification; the gDNA removal step may be omitted when DNA-free RNA is used. Guaranteed specificity and detection rates for PCR and RT-qPCR applications. |
| Reaction Conditions / Protocol | Reverse transcription is completed in 5 minutes (total workflow under 6 minutes). For direct synthesis of fragments longer than 10 kb, extend the reverse transcription time to 20–30 minutes to ensure full-length product recovery. Choose Random Primers N6, Oligo(dT)18, or gene-specific primers as the reverse transcription primer according to the experimental design. |
| Components / Formulation | 4× AdvanceFast SuperMix (with blue tracking dye) 5× gDNA Digester Mix Random Primers (50 μM) Oligo(dT)18 Primers (50 μM) RNase-free Water |
| Storage Conditions | Store at -20°C. The reverse transcription product can be used immediately in downstream reactions or stored short-term at -20°C; for long-term storage, aliquot and keep at -80°C, avoiding repeated freeze-thaw cycles. |
| Shelf Life | 12 months when stored at -20°C. |
| Package Specifications | 10 T 100 T |
| Product Form | Kit |
| Quality Control | Reverse transcription efficiency benchmarked against leading competing products: 5-minute reverse transcription yields Ct values equal to or lower than competitors using 13- or 20-minute protocols. Addition of the tracking dye verified to have no adverse effect on Ct values, melt-curve peaks, or Tm values. |
| Expiration Date / Stability | Valid for 12 months at -20°C. The kit remains stable for 14 days at 4°C; the cDNA product remains stable for 7 days at 37°C and withstands 50 freeze-thaw cycles. |
| Compatibility | The single-stranded cDNA product can be used directly in downstream PCR or qPCR reactions and is compatible with standard qPCR reagents. The kit does not contain RNase H. A dye-free version without the blue tracking dye is also available for applications where the tracer may be a concern. |
| Application Notes / Precautions | The blue tracking dye indicates pipetting progress and has been verified not to affect reverse transcription efficiency or downstream qPCR Ct values, melt-curve peaks, or Tm values; a dye-free version is available for tracer-sensitive applications. The kit contains no RNase H. For fragments longer than 10 kb, extend the reverse transcription time to 20–30 minutes to ensure recovery of full-length cDNA. For long-term storage of the cDNA product, aliquot and store at -80°C; avoid repeated freeze-thaw cycles. |
For research use only, not for clinical use.
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