"
Team:SYSU-China/file/Project/Notebook/Protocol/Mutation.html
From 2014.igem.org
(Difference between revisions)
Shenjian412 (Talk | contribs) |
|||
| Line 1: | Line 1: | ||
<!--SYSUCHINA--> | <!--SYSUCHINA--> | ||
<h1>Mutation</h1> | <h1>Mutation</h1> | ||
| + | |||
| + | <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> | ||
| + | |||
| + | |||
| + | |||
<!--SYSUCHINA!--> | <!--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.
