Author:
Hung Jui-Hung,Weng Zhiping
Abstract
Designing oligonucleotide primers is a crucial step for successful molecular biology experiments that require the use of the polymerase chain reaction (PCR). PCR involves cycles of three steps: denaturation, annealing, and extension. During denaturation, double-stranded DNA (dsDNA) molecules (templates) are separated into single strands. During annealing, a pair of primers is annealed to the complementary regions of the single-stranded molecules. In the extension step, DNA polymerase extends the primers to produce DNA molecules that correspond to the region bracketed by the primers (the amplicons). All of these steps are temperature sensitive, and the common choice of temperatures is 94°C, 60°C, and 70°C, respectively. Poorly designed primers may lead to no amplification product or additional undesired amplified fragments. The goals of primer design include good primer specificity, high annealing efficiency, appropriate melting temperature, proper GC content, and the prevention of primer hairpins or primer dimers.
Publisher
Cold Spring Harbor Laboratory
Subject
General Biochemistry, Genetics and Molecular Biology
Reference11 articles.
1. QuantPrime – a flexible tool for reliable high-throughput primer design for quantitative PCR
2. Burpo FJ . 2001. A critical review of PCR primer design algorithms and cross hybridization case study. Computational Biology Department, Biochemistry 218, Stanford University, California ( http://biochem218.stanford.edu/Projects2001/Burpo.pdf ).
3. GeneFisher—Software support for the detection of postulated genes;Proc Int Conf Intell Syst Mol Biol,1996
4. OSP: a computer program for choosing PCR and DNA sequencing primers.
5. A computer program for selection of oligonucleotide primers for polymerase chain reactions
Cited by
18 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献