白斑综合征病毒PCR检测中使用dUTP扩增的PCR产物极限长度

摘要

To prevent carry-over contamination, dTTP was replaced by dUTP in PCR detection for white spot syndrome virus (WSSV). Seven pairs of primers were designed according to the genome sequence of WSSV. The 600~l 800bp DNA fragments were amplified separately. It was found that the utmost length of PCR product is about 1 400bp, when dUTP is used to take place of dTTP.%依据白斑综合征病毒的序列设计了7对PCR引物,扩增长度从600bp到1 800bp.结果显示,使用dUTP代替dTTP,用普通Taq酶所能扩增的最大长度约为1 400bp.Mg2+梯度研究发现,扩增片段长度越长,最适Mg2+浓度越低.使用具有3,-5’外切酶活性的Taq酶无法扩增出任何长度的片段.