PCR快速筛选重组克隆方法的建立  

Rapid Screening of Recombinant Clone by PCR

作　　者：张传生[1] 耿立英[2] 杨娜娜[2] 孟春花[2] 朱靖[2] 杜立新[3] 

机构地区：[1]山东农业大学生命科学学院,山东泰安271018 [2]山东农业大学动物科技学院,山东泰安271018 [3]中国农业科学院畜牧研究所,北京100094

出　　处：《生物技术》2005年第1期43-44,共2页Biotechnology

基　　金：国家自然科学基金项目 (30 2 70 949) ;山东省自然科学基金项目 (编号 :Y97D0 30 62 )资助 ~~

摘　　要：目的 :建立一种PCR技术快速筛选重组克隆的方法。方法 :用Triton裂解菌落作为模板 ,以pMD - 18T载体多克隆位点设计的引物与目的基因猪 β-INF引物设计不同组合 ,PCR扩增待检重组克隆。 结果 :PCR技术快速筛选出猪 β -INF重组克隆并鉴定了插入方向。结论To set up a new method for rapid screening of recombinant clone.By taking the sequences from the plasmid vector and sequences from the inserted gene as the primer pairs,a method has been constructed with 5% Triton X-100 lysis buffer and PCR for scalping and identifying positive recombination plasmids conveniently and fast.The positive clone and orientation of DNA insertion were conveniently determined and the results were in good accordance with those by traditional restriction enzyme methods.This method could screen recombinant clone rapidly,while identifying the orientation and length of the inserted sequence.

关 键 词：重组克隆 筛选 PCR 

分 类 号：S813.13[农业科学—畜牧学]