Abstract
Quantitative real-time polymerase chain reaction (qPCR) is a powerful tool for analysis and quantification of gene expression. It is advantageous compared to traditional gel-based method of PCR, as gene expression can be visualized “real-time” using a computer. In qPCR, a reporter dye system is used which intercalates with DNA’s region of interest and detects DNA amplification. Some of the popular reporter systems used in qPCR are the following: Molecular Beacon®, SYBR Green®, and Taqman®. However, success of qPCR depends on the optimal primers used. Some of the considerations for primer design are the following: GC content, primer self-dimer, or secondary structure formation. Freely available software could be used for ideal qPCR primer design. Here we have shown how to use some freely available web-based software programs (such as Primerquest®, Unafold®, and Beacon designer®) to design qPCR primers.
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Thornton, B., Basu, C. (2015). Rapid and Simple Method of qPCR Primer Design. 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_13
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DOI: https://doi.org/10.1007/978-1-4939-2365-6_13
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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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