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| Product Name | AdvanceFast 1st Strand cDNA Synthesis SuperMix for qPCR |
| Catalog No. | RESK-0020 |
| Description | The AdvanceFast 1st Strand cDNA Synthesis SuperMix for qPCR is an upgraded, ready-to-use reverse transcription premix that offers stronger tolerance to residual inhibitors, faster reaction speed, and higher cDNA yields than the previous generation of reverse transcriptases. The SuperMix contains every component required for reverse transcription and tolerates reaction temperatures up to 65°C, making it well suited for RNA templates with complex secondary structures. Only RNA template and RNase-free water need to be added to start the reaction. The resulting cDNA is intended for qPCR and is not recommended for long-fragment applications such as gene cloning; for those needs, Alta DiagnoTech's AdvanceFast 1st Strand cDNA Synthesis Kit is recommended. |
| Intended Use | First-strand cDNA synthesis from RNA templates for downstream qPCR analysis. Not intended for long-fragment amplification such as gene cloning. |
| Principle / Technology | Ready-to-use reverse transcription premix containing an engineered reverse transcriptase with enhanced inhibitor tolerance and thermostability (up to 65°C). A defined ratio of Random Primers N6 and Oligo dT is pre-mixed into the SuperMix, so no additional primers are required for routine RNA qPCR workflows. |
| Detection Method | Reverse transcription followed by dye-based qPCR quantification. |
| Sample Type | Total RNA from various species and targets with different GC contents, including degraded RNA and RNA containing inhibitors; compatible with 10 pg to 5 μg RNA input. |
| Performance Range / Specifications | Linear cDNA synthesis over a 10 pg to 1 μg RNA input range with correlation coefficient R2 up to 0.99; reverse transcription efficiency up to 96.4% (293T cell RNA) and 100% (Arabidopsis RNA); tolerates reaction temperatures up to 65°C. |
| Sensitivity / LOD | Detects RNA input as low as 10 pg. |
| Specificity | Higher cDNA synthesis efficiency and lower Ct values across targets of different species and GC contents compared with other reverse transcription reagents. |
| Reaction Conditions / Protocol | Tolerates reverse transcription temperatures up to 65°C. After the reaction, inactivate at 85°C, then hold at 4°C. Extended reverse transcription times are not recommended, as they may delay Ct values; follow the program specified in the manual. |
| Components / Formulation | AdvanceFast SuperMix: 60 μL (10 T) / 600 μL (100 T) RNase-free Water: 200 μL (10 T) / 1 mL x 2 (100 T) |
| Storage Conditions | Store at -25°C to -15°C. Once opened, the reagent may be kept at 4°C provided it is free from pipette-tip cross-contamination, but use within 2 weeks is recommended. |
| Shelf Life | 12 months when stored at -25°C to -15°C. |
| Package Specifications | 10 T, 100 T |
| Product Form | Premix (SuperMix) |
| Quality Control | cDNA products remain stable at room temperature for 7 days; reagents withstand 30 repeated freeze-thaw cycles. |
| Expiration Date / Stability | Valid for 12 months at -25°C to -15°C; cDNA product stable at room temperature for 7 days; reagent stable through 30 freeze-thaw cycles. |
| Compatibility | Compatible with 10 pg to 5 μg RNA input. For downstream qPCR, Alta DiagnoTech's dye-based qPCR reagents are recommended. For small RNA, circular RNA, or long-fragment PCR, use a kit with separate primers such as Alta DiagnoTech's 1st Strand cDNA Synthesis Kit III (with gDNA Digester) or AdvanceFast 1st Strand cDNA Synthesis Kit. |
| Application Notes / Precautions | Random Primers N6 and Oligo dT are pre-mixed in the SuperMix; no additional primers are needed for routine RNA qPCR. Mix reagents by gentle inversion followed by brief centrifugation; do not vortex vigorously. Excessive RNA input may introduce inhibitory carryover that affects downstream qPCR; dilute the cDNA if Ct values increase or amplification curves become erratic. Do not extend the reverse transcription time beyond the recommended program, as this may delay Ct values. After 85°C inactivation, set the thermal cycler to hold at 4°C to prevent heat damage to products left in the instrument. No concentration measurement or adjustment of the RT product is required before qPCR; dNTPs and other residual components interfere with spectrophotometric readings. The cDNA product can be left at room temperature for up to one week without affecting subsequent experiments. |
For research use only, not for clinical use.
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