Yu Bing, Zhang Changbin
Department of Molecular and Clinical Genetics, Royal Prince Alfred Hospital, and Sydney Medical School, Central, The University of Sydney, Camperdown, NSW, Australia.
Methods Mol Biol. 2011;760:91-107. doi: 10.1007/978-1-61779-176-5_6.
In silico PCR analysis is a useful and efficient complementary method to ensure primer specificity for an extensive range of PCR applications from gene discovery, molecular diagnosis, and pathogen detection to forensic DNA typing. In silico PCR, SNPCheck, and Primer-BLAST are commonly used web-based in silico PCR tools. Their applications are discussed here in stepwise detail along with several examples, which aim to make it easier for the intended users to apply the tools. This virtual PCR method can assist in the selection of newly designed primers, identify potential mismatches in the primer binding sites due to known SNPs, and avoid the amplification of unwanted amplicons so that potential problems can be prevented before any "wet bench" experiment.
电子PCR分析是一种有用且高效的补充方法,可确保引物特异性,适用于从基因发现、分子诊断、病原体检测到法医DNA分型等广泛的PCR应用。电子PCR、SNPCheck和Primer-BLAST是常用的基于网络的电子PCR工具。本文将逐步详细讨论它们的应用,并给出几个例子,旨在使目标用户更容易应用这些工具。这种虚拟PCR方法可以帮助选择新设计的引物,识别由于已知单核苷酸多态性(SNP)导致的引物结合位点潜在错配,并避免扩增不需要的扩增子,从而在任何“湿实验室”实验之前预防潜在问题。