微波介导的载体快速转化及克隆筛选方法

Accelerated bacterial transformation and, recombinant screening by microwave radiations method

应用微波辐射进行载体快速转化及重组体鉴定。经选择辐射转化条件,表明在低渗钙离子(0.05mol/L或0.1mol/L)条件下,150W、45s的辐射可得到理想的转化率。采用微波碎菌抽提质粒方法对重组体进行快速鉴定,不需特殊设备,省时、可靠。用该方法对肾综合征出血热病原汉坦病毒76-118株S片段基因进行克隆、转化和重组体鉴定试验,证实其具有简易、快速、高效等优点。

We reported the mothed for transformation and screening rapidly using microwave radiations. The optimal choice of the protocal depended on the verification of several variable or conditions. We recommended the microwave radiations for 45 s at power 150 W that obtain a maximum number of transformants. Moreover, we described a novel modification of microwave-lysis plasnnd DNA preparation method that deceased the time and labor requirements of the procedure while increasing its reliability. The S segment gene of HFRSV Serotype HTN 76-118 was cloned and the re-combinants were screened using microwave radiations. It proved the method was simple, easy and rapid.