Primer Spanner:一个高效的定点突变PCR引物设计在线工具

Primer Spanner: A Web- based Platform to Design PCR Primers for High Efficient Site-directed Mutagenesis

基于PCR的实验策略在生物工程研究中具有广泛应用,如定点突变(site-directed mutagenesis,SDM),DNA拼接和载体构建。引物设计是这类实验技术中的关键一环,因其直接影响扩增效率和PCR产物的拼合。在嵌合式引物设计方法(一对突变引物在5'端具有互补序列)的基础上,开发了一个在线工具Primer Spanner(PS),可简单高效获得设计定点突变引物。PS可应用于单碱基或连续多碱基替换、插入、敲除等突变形式。通过大量突变实验与测序验证,结果表明该工具设计的引物进行的定点突变效果良好1)。

PCR-based strategy provides economical and efficient application in site-directed mutagenesis （SDM） and DNA assembling such as PCR based vector construction. In these processes, primers are extremely important as they determing amplification efficiency of the target DNA fragments, also influencing circularization of the new plasmids. So this web-based software Primer Spanner （PS） was developed to design primer pairs with partially complementary 5＇-ends, which is a special tool for chimeric primer designing. Primers designed by this tool are applied to substitution, insertion and deletion for single-site or multi-site mutagenesis. The performance of the software had been verified by designed experiment and DNA sequencing. The PS primer designing engine is freely accessible at ： http ：//PS1. biocloud, org. cn.