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Primer Design Using Primer Express® for SYBR Green-Based Quantitative PCR

  • Amarjeet Singh
  • Girdhar K. PandeyEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1275)

Abstract

To quantitate the gene expression, real-time RT-PCR or quantitative PCR (qPCR) is one of the most sensitive, reliable, and commonly used methods in molecular biology. The reliability and success of a real-time PCR assay depend on the optimal experiment design. Primers are the most important constituents of real-time PCR experiments such as in SYBR Green-based detection assays. Designing of an appropriate and specific primer pair is extremely crucial for correct estimation of transcript abundance of any gene in a given sample. Here, we are presenting a quick, easy, and reliable method for designing target-specific primers using Primer Express® software for real-time PCR (qPCR) experiments.

Key words

Real-time PCR SYBR Green Primer Expression Primer Express® 

Notes

Acknowledgement

Research work in GKP’s lab is partially supported by grants from University of Delhi, Department of Biotechnology (DBT), Department of Science and Technology (DST), and Council of Scientific and Industrial Research (CSIR), India. AS acknowledges CSIR for research fellowship.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Plant Molecular BiologyUniversity of Delhi South CampusNew DelhiIndia

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