重组克隆PCR扩增快速筛选方法的建立

Rapid screening of recombinant clone by PCR

目的 :建立PCR扩增快速筛选重组克隆的方法。方法 :利用载体本身的启动子序列和目的片段的特异性引物作为引物 ,通过合理搭配 ,用常规PCR方法扩增待检重组克隆。结果 :可以方便地筛选出重组克隆以及顺向和反向重组克隆 ,与传统的酶切鉴定结果一致。结论 :该方法可以快速、准确的筛选重组克隆 ,并能够确定片段插入的方向和大小。

Aim:To set up a new method for rapid screening of recombinant clone.Methods:By taking the DNA sequences from the plasmid vector and sequences from the inserted gene as the primer pairs, a RCR based recombinant clone screening method was established.Results: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.Conclusion:This method could screen recombinant clone rapidly, while identifying the orientation and length of the inserted sequence.