Then the pcr product was purified , ligated into pgem - t vector by ta cloning 筛选阳性克隆,测序,大量制备序列完全正确的质粒。
The pcr product was cloned into pmd18 - t vector . the positive recombinant clone was identified by pcr and endonuclease digest 提取重组质粒经pcr鉴定和酶切鉴定后,对插入片段进行序列测定及分析。
The product of pcr named vp6 is approximate 1 . 3kb in length . the vp6 gene was cloned into pmd18 - t vector and sequenced 将其插入克隆载体质粒pmd18 - t的ecorv酶切位点处,构建重组质粒pmd18 - t ? vp6 。
Pgem - t vector system , jm109 competent cells , reverse transcription kit and in vitro translation kit were purchased from promega company Pgem一t载体,感受态细菌jm109 ,反转录试剂盒和体外翻译试剂盒均购自promega公司。
2 . pcr products can be purified by ctab method , which removing the dntp and primers . the purified products can be used in the ligation reaction to pmd18 - t vector with high efficiency 2 、采用ctab进行pcr产物的纯化,以去除pcr产物中的引物和dntp ,所得产物可用于与t载体连接反应,以提高连接效率。