PCR快速筛选重组克隆方法的建立

Rapid Screening of Recombinant Clone by PCR

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.