聚合酶链反应中引物的计算机辅助筛选

A COMPUTER-AIDED SELECTION OF OLIGONUCLEOTIDE PRIMER FOR POLYMERASE CHAIN REACTION

利用计算机辅助筛选聚合酶链反应（ＰＣＲ）的扩增引物。计算机程序根据引物筛选规则编写。这些规则包括：①引物的３′端均以ＧＣ对结尾；②引物的长度为１８～的个寡核苷酸；③每个引物中的ＧＣ含量为４５％～６０％；④避免引物自身及引物之间的同源性。由此我们设计了ＳＲＳ小鼠白血病病毒的引物，成功地扩增了３．４ｋｂ的大片段目的基因。实验结果证明了由此法筛选出来的引物的有效性和特异性。

We have designed a computer program which can scan nucleic acid sequences to select primers for polymerase chain reaction.This program is based on a set of rules for selection primers.These rules include:(1)Primers should contain a GCtype sequence pair at their 3'end;(2)The length of each primer should be between 18 to 24 nucleotides;(3) Each primer should have a GC-type sequence content of between 45% to 60% of its total bases;(4)Limit primer loss occurring from primer-self or primer-primer homologies.We have designed a pair of primers of SRS leukemia virus by this method,and successfully amplified 3.4kb products of interest.The results satisfied the effectiveness and specificity of primers selected by this method.