Transformation of plasmid DNA into E. coli using the heat shock method.
Transformation of plasmid DNA into E. coli using the heat shock method.
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DOI：
10.3791/253
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发表时间：
2007-01-01
期刊：
影响因子：
--
通讯作者：
Hall, James E
中科院分区：
文献类型：
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作者：
Froger, Alexandrine;Hall, James E
利用热休克方法将质粒DNA转化到大肠杆菌是分子生物学的一项基本技术。它包括将外源质粒或连接产物插入细菌中。这个视频协议描述了使用Genlantis的商业上可以买到的化学活性细菌进行转化的传统方法。在冰中孵育一小段时间后，将具有化学活性的细菌和DNA的混合物置于42摄氏度的45秒(热休克)中，然后再放回冰中。加入SOC培养液，将转化细胞在37℃下搅拌培养30分钟。为了确保无论转化效率如何都能分离出菌落，两个数量的转化细菌被平板化。这一传统的方案可以成功地用于转化大多数商业上可用的可竞争细菌。Genlantis的涡轮电池还可以用于说明书中描述的新型3分钟转换协议。
Transformation of plasmid DNA into E. coli using the heat shock method is a basic technique of molecular biology. It consists of inserting a foreign plasmid or ligation product into bacteria. This video protocol describes the traditional method of transformation using commercially available chemically competent bacteria from Genlantis. After a short incubation in ice, a mixture of chemically competent bacteria and DNA is placed at 42 degrees C for 45 seconds (heat shock) and then placed back in ice. SOC media is added and the transformed cells are incubated at 37 degrees C for 30 min with agitation. To be assured of isolating colonies irrespective of transformation efficiency, two quantities of transformed bacteria are plated. This traditional protocol can be used successfully to transform most commercially available competent bacteria. The turbocells from Genlantis can also be used in a novel 3-minute transformation protocol, described in the instruction manual.