Single Gene qPCR Primer Set

Single Gene qPCR Primer Set is a service to verify the primers of dsDNA binding dye type used for real-time PCR analysis according to MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) guideline *. Gene information to be analyzed will be provided as a primer set according to the MIQE guideline * and the target specificity and PCR efficiency will be verified and provided as a primer set of 200rxn. We also provide verification services for the retention primer sequence. It can be used directly in the experiment without any further verification, and you can derive ready-to-publish data. * The MIQE Guidelines [Bustin, S.A., et al. 2009. The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments, Clinical Chemistry 55:4, 611-622]

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Cat. No.
S-6042-S200

Overview

Single Gene qPCR Primer Set is a service that provides a primer set after performing a verification test to check whether it meets the required condition according to the MIQE guideline *. It can be easily used for experiments without any further verification and can save time for primer optimization. (Table References)

*MIQE Guideline?

MIQE (Minimum Information for Publication of Quantitative Real-time PCR Experiments) Guideline is for evaluating the reliability and reproducibility of qPCR experiment providing step-by-step checklists. This service provides the experimental information to the customers for them to evaluate the validity of the results.

Table. Information required for qPCR validation according to the MIQE guideline

Item to check
Specificity (gel, sequence, melt, or digest)
For dsDNA binding dyes, Cq of the NTC
Calibration curves with slope and y intercept
PCR efficiency calculated from slope
r2 of calibration curve
Linear dynamic range
Cq variation at LOD
Evidence for LOD

The MIQE Guidelines [Bustin, S.A., et al. 2009. The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments, Clinical Chemistry 55:4, 611-622]

Features and Benefits

  • Target-specific primer design using primer blast and our bioinformatics tool

  • Exclusion of self primer-dimer formation sequence 

  • Identification of single peak in the dissociation curve

  • Identification of single band in Gel electrophoresis

  • Short amplicon size of 70 ~ 150 bp

  • Wide amplification range of copies being about 102 ~ 107 

  • qPCR amplification efficiency of 90-110% in compliance with the MIQE Guideline

Application

• Applicable to Quantitative Real-time PCR

Experimental data

Figure. A diagram showing dissociation curve, dynamic range, standard curve, and electrophoresis image of the human ACTB and its target.

* As Single Gene qPCR Primer Set is verified with ExiCycler™ 96 V3 and 2X GreenStar™ qPCR Master Mix (-ROX Dye)(K-6253), results may be different if other company products are used.

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