qScript cDNA Synthesis Kit
Features & Benefits
- Economical 2-component kit ideally suited for high throughput expression studies
- Optimized, double pre-primed master mix component ensures equal representation of 5' and 3' RNA sequences ≤ 1kb for quantitative and conventional two-step RT-PCR
- Broad linear dynamic detection range with limiting (10pg) or plentiful (1ug) samples of total RNA
qScript cDNA Synthesis Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
The qScript cDNA Synthesis Kit is a sensitive and easy-to-use solution for RNA quantification using two step RT-qPCR. The novel qScript Reaction Mix provides all the necessary components for cDNA synthesis except enzyme and RNA template. qScript reverse transcriptase is a mixture of an engineered MMLV RT and a ribonuclease inhibitor protein. This economical, highly stable stabilize, 2-component reagent system has been rigorously optimized to ensure sensitive and linear RNA detection with a wide-range of input RNA and relative abundance. Reagent performance is unaffected by repetitive freeze/thaw cycling (>20X), conferring greater ease-of-use and assay performance consistency. Oligo (dT) and random primers are pre-blended in a precise ratio to provide equal representation of 5' and 3'-sequences for accurate gene expression quantification. The resulting cDNA product is directly compatible with all qPCR chemistries or conventional end-point RT-PCR of amplified fragments ≤1 kb in length.
For gene-specific priming (GSP) or two-step RT-PCR of RNA exceeding 1kb in total length, see our qScript® Flex cDNA Kit.
RT-qPCR Dynamic Range
Broad Linear Dynamic Range
qScript cDNA Synthesis Kit was used for first-strand cDNA synthesis using log-fold serial dilutions of HeLa cell total RNA from 1 µg to 1 pg. Six replicate cDNA reactions were performed for each input quantity of RNA. 1/10th of each first-strand reaction was used for qPCR of the b-actin gene using PerfeCTa™ SYBR Green SuperMix.
Consistent cDNA Synthesis Efficiency - cDNA serial dilution vs. total RNA serial dilution
qPCR Standard Curve (Orange): Log fold serial dilutions of qScript-synthesized cDNA from 1 µg of HeLa cell total RNA were used as template for qPCR of ACTB using PerfeCTa™ SYBR Green SuperMix. 2-Step qRT-PCR Standard Curve (Blue): qScript cDNA Synthesis Kit was used for first-strand cDNA synthesis using log-fold serial dilutions of HeLa cell total RNA from 1 µg to 1 pg. Six replicate cDNA reactions were performed for each input quantity of RNA. 1/10th of each first-strand reaction was used for qPCR of the ACTB gene using PerfeCTa™ SYBR Green SuperMix. Concordance of the two standard curves is indicative of consistent conversion of RNA into cDNA with high efficiency at each amount of RNA tested. qScript cDNA Synthesis Kit enables reliable quantification of RNAs from low amounts of starting material.
- 5X concentrated master mix containing: titered primer blend (oligo dT(20) and random hexamer), qPCR-optimized dNTP blend and flexible magnesium titration
- 20X concentrated qScript reverse transcriptase
- Nuclease-free water
Storage & HandlingStore components in a constant temperature freezer at -25°C to -15°C upon receipt. After thawing, mix thoroughly before using. For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.
Thaw completely on ice then pulse vortex to mix and briefly spin-down to collect contents before opening tube. Concentrated reagents are viscous. When drawing out material, touch pipette tip to liquid interface but do not immerse.