Team:USTC CHINA/Notebook/Protocols/The transformation of Bacillus subtilis

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<h1>The transformation of Bacillus subtilis</h1>
<p>- thaw one aliquot at 37 °C</br>
- use these cells to inoculate 20 ml LS medium</br>
- shake cells slowly in a 30°C water bath to obtain maximal competence (about 2 h)</br>
- take 1 ml aliquots into a glass tube or 2 ml Eppendorf tube, add 10 μl of 0.1 M
EGTA (CB-0732-10GAM), and incubate for 5 min at room temperature</br>
- add plasmid or chromosomal DNA and incubate for 2 h at 37°C while well shaking
(well mixing is important when using Eppendorf cups)</br>
- if glass tubes were used, transfer cell suspension into an Eppendorf tube</br>
- centrifuge, discard supernatant carefully and resuspend the cells into the final
supernatant remaining on the pellet</br>
- plate on selective 2xYT medium</br>
- incubate at 37°C overnight</br>


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