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| Product Name | 1st Strand cDNA Synthesis SuperMix II for qPCR (with gDNA Digester) |
| Catalog No. | RESK-0026 |
| Description | This ready-to-use premix is developed on our second-generation thermostable reverse transcriptase and is optimized for two-step RT-qPCR. The enzyme offers greatly improved thermostability, tolerating reaction temperatures up to 50°C, making it suitable for reverse transcribing RNA templates with complex secondary structures, and its enhanced template affinity supports low-input templates and low-copy-number genes. The kit includes a gDNA digester that removes residual genomic DNA contamination from RNA templates for more reliable downstream results, together with a 2x SuperMix plus containing every component required for reverse transcription (buffer, dNTPs, reverse transcriptase, RNase inhibitor, and a Random Primers/Oligo (dT)18 primer mix). Only template RNA and RNase-free ddH2O need to be added; the reverse transcription step simultaneously terminates gDNA digester activity, protecting cDNA integrity. The Random Primers/Oligo (dT)18 ratio is optimized for qPCR so that cDNA synthesis initiates uniformly across all regions of RNA transcripts with equal efficiency, maximizing the accuracy and reproducibility of qPCR results. |
| Intended Use | First-strand cDNA synthesis with genomic DNA removal for two-step RT-qPCR-based gene expression analysis. For research use. |
| Principle / Technology | Based on a second-generation thermostable reverse transcriptase that tolerates reaction temperatures up to 50°C and shows enhanced template affinity, enabling reverse transcription of RNA templates with complex secondary structures and of low-input or low-copy-number targets. A supplied gDNA digester eliminates residual genomic DNA contamination from RNA templates, and its activity is terminated as the reverse transcription reaction proceeds, ensuring cDNA integrity. The Random Primers/Oligo (dT)18 ratio is optimized for qPCR so cDNA synthesis initiates from all regions of RNA transcripts with equal reverse transcription efficiency. |
| Detection Method | Downstream qPCR; the reverse transcription product is compatible with both SYBR Green and probe-based (TaqMan) qPCR. |
| Sample Type | Template RNA (total RNA in the range of 1 ng to 5 ug for routine expression-level transcripts). |
| Performance Range / Specifications | Reverse transcription reactions can be performed at temperatures up to 50°C; good reproducibility. |
| Sensitivity / LOD | Enhanced template affinity makes the product suitable for low-input templates and low-copy-number genes; applicable to total RNA inputs of 1 ng to 5 ug. |
| Specificity | The qPCR-optimized Random Primers/Oligo (dT)18 primer ratio enables cDNA synthesis to initiate from all regions of RNA transcripts with equal reverse transcription efficiency, maximizing the authenticity and reproducibility of qPCR results. |
| Reaction Conditions / Protocol | Add template RNA and RNase-free ddH2O to the premix to run the reverse transcription reaction; reaction temperatures up to 50°C can be applied. The reverse transcription step simultaneously terminates gDNA digester activity. It is recommended to dilute the cDNA product 5- to 10-fold before qPCR. |
| Components / Formulation | gDNA digester 2x cDNA Synthesis SuperMix plus (contains reaction buffer, dNTPs, second-generation reverse transcriptase, RNase inhibitor, and Random Primers/Oligo (dT)18 primer mix) |
| Storage Conditions | Store at -20°C. |
| Shelf Life | 18 months. |
| Package Specifications | 10 T; 100 T |
| Product Form | Ready-to-use premix (SuperMix) |
| Source / Origin | Developed on a second-generation thermostable reverse transcriptase. |
| Shipping Conditions | Shipped on dry ice. |
| Expiration Date / Stability | Stable for 18 months when stored at -20°C. |
| Compatibility | The reverse transcription product is compatible with SYBR Green and probe-based qPCR and can be combined with Alta DiagnoTech qPCR SYBR Green Master Mix or qPCR TaqMan Probe Master Mix for high-performance gene expression analysis. After dilution, the cDNA product can also be used with qPCR reagents from other suppliers. |
| Application Notes / Precautions | The premix uses Random Primers and Oligo (dT)18 premixed at an optimized ratio for qPCR. The cDNA product is not recommended for downstream conventional PCR: the mixed primers produce uniform but relatively short cDNA, suitable only for short amplicons. For downstream PCR, use a kit-format product with Oligo (dT) primers instead. Dilute the reverse transcription product 5- to 10-fold before qPCR to reduce pipetting error, improve reproducibility, and minimize inhibition of the qPCR reaction by concentrated reverse transcription components. After dilution, the product is compatible with qPCR reagents from other brands. |
| Batch-to-Batch Consistency | Good reproducibility. |
For research use only, not for clinical use.
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