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.
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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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Singh, A., Pandey, G.K. (2015). Primer Design Using Primer Express® for SYBR Green-Based Quantitative PCR. In: Basu, C. (eds) PCR Primer Design. Methods in Molecular Biology, vol 1275. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2365-6_11
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DOI: https://doi.org/10.1007/978-1-4939-2365-6_11
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2364-9
Online ISBN: 978-1-4939-2365-6
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