qScript™ cDNA Synthesis Kit
qScript cDNA Synthesis kits – for exceptional cDNA synthesis and accurate representation from less starting material.
The qScript™ cDNA Synthesis Kit is a sensitive and easy-to-use solution for RNA quantification using two step RT-PCR. This kit contains an optimised blend of random and oligo (dT) primers for robust, consistent and unbiased first-strand synthesis over a broad range of RNA template concentrations. The novel qScript Reaction Mix (5X) contains all required reagents for cDNA synthesis, except for qScript Reverse Transcriptase and RNA. qScript Reverse Transcriptase is provided in a separate tube.
qScript cDNA SuperMix is a sensitive and easy-to-use 1-tube reagent for first-strand cDNA synthesis that combines a highly-modified RNAse H+ mutant of M-MLV together with ribonuclease inhibitor protein (RIP) in a rigorously optimized formulation for real-time qPCR applications. The stabilized 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. For gene-specific priming (GSP) or two-step RT-PCR of RNA exceeding 1kb in total length, see our qScript® Flex cDNA 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 (4ug) samples of total RNA
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 Efficiency:
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.
Contents
- 5X concentrated master mix containing: titered primer blend (oligo dT(20) and random hexamer), qPCR-optimized dNTP blend and flexible magnesium titration
- 50X concentrated qScript reverse transcriptase
- Nuclease-free water
Storage conditions
qScript cDNA Synthesis Kit components are stable for up to 1 year when stored in a constant temperature freezer at or below -20°C. Reagent performance is unaffected by 20+ repetitive freeze-thaw. 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.
Resources
Ordering
| Cat# 95047-025 | 25 rxns | |
| Cat# 95047-100 | 100 rxns | |
| Cat# 95047-500 | 500 rxns |