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Team:SYSU-China/file/Project/Notebook/Protocol/Mutation.html
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<h1>Mutation</h1> | <h1>Mutation</h1> | ||
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| + | <h2><b>Test the mutation rate</b></h2> | ||
| + | <p>1. Co-transform the mutagenic plasmid and reporter plasmid (RFP) into BL21. </p> | ||
| + | <p>2. Pick the red clones and add in Amp+ plus Cm+ LB, add IPTG, culture in 37℃ overnight. </p> | ||
| + | <p>3. Dilute the bacterium solution and spread it on LB plate. Incubate overnight, dilute the rest liquid LB to 1:1000 with new one and continue to culture. </p> | ||
| + | <p>Count and pick the white clones, confirm the mutation by DNA sequencing. </p> | ||
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| + | <!--SYSUCHINA!--> | ||
Latest revision as of 02:09, 18 October 2014
Mutation
Test the mutation rate
1. Co-transform the mutagenic plasmid and reporter plasmid (RFP) into BL21.
2. Pick the red clones and add in Amp+ plus Cm+ LB, add IPTG, culture in 37℃ overnight.
3. Dilute the bacterium solution and spread it on LB plate. Incubate overnight, dilute the rest liquid LB to 1:1000 with new one and continue to culture.
Count and pick the white clones, confirm the mutation by DNA sequencing.
