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qScript cDNA Synthesis Kit

Economical 2-component kit ideally suited for high throughput gene-expression studies
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.

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qScript cDNA Synthesis Kit

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Kit Size:  25 x 20 μL rxns (1 x 25 µL)
Part Number:  95047-025
Price:  $130.00
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Kit Size:  100 x 20 μL rxns (1 x 100 µL)
Part Number:  95047-100
Price:  $322.00
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Kit Size:  500 x 20 μL rxns (1 x 500 µL)
Part Number:  95047-500
Price:  $1,353.00
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Description

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.

Details

Details

Contents
  • 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

Details

Contents
  • 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

Performance Data

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CofA (PSFs)

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SDSs

FAQs

Does the qScript™ cDNA Synthesis Kit include an RNase Inhibitor like the qScript™ cDNA SuperMix?
Both kits contain an RNase inhibitor protein. qScript™ cDNA SuperMix is provided as a single tube reaction of 5X concentrated master mix. It provides all necessary components for first-strand synthesis including: buffer, dNTPs, MgCl2, primers, RNase inhibitor protein, qScript reverse transcriptase and stabilizers (except RNA template) . In the case of the qScript™ cDNA Synthesis Kit, Rnase inhibitor is premixed with the reverse transcriptase and is provided as a concentrated enzyme.
For first strand cDNA synthesis, is it better to use oligo(dT), random hexamers, gene specific primer (GSP) or combination of these primers?
The choice of primer depends on your experimental goals. Oligo(dT) is recommended when using total RNA for cDNA synthesis. It is the key to full-length cDNA synthesis. Random hexamers give a series of short first-strand products spanning the entire mRNA. Use of random hexamers may be helpful if the PCR fragment is at the 5´ end of a large mRNA. To ensure full-length cDNA synthesis of large transcripts, oligo(dT) can be added along with random hexamers during first-strand synthesis. Gene specific primers (GSP) for cDNA synthesis may also be used and are required in a few applications such as 5´ RACE and qRT-PCR. For GC-rich templates, or templates rich in secondary structure, a GSP may not work as well as priming with oligo dT for first strand synthesis. If an RT-PCR is problematic, trying different options of oligo dT, random primers and/or GSP for priming first strand synthesis may find a solution. Oligo(dT)20 primer (Cat. No. 18418-020) is recommended for use with SuperScript III Reverse Transcriptase
What is the smallest quantity of RNA detectable by the qScript™ cDNA Synthesis Kit?
Lower limits of detection by the qScript™ cDNA Synthesis Kit are dependent on many factors, such as primer design, target size, and the abundance of message. However, this system was able to detect GAPDH mRNA from as little as 1.0 pg of total HeLa RNA when used in conjunction with AccuStart Taq DNA Polymerase.
What is the RNA input for the qScript cDNA Synthesis Kit?
The qScript cDNA Synthesis Kit provides for the quantitative conversion of 1µg to 10 pg total RNA to cDNA, with a reaction volume of 20 ul.

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