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Team:Elan Vital South Korea/l transformation
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| - | <p> | + | <p>a. Take out 50㎕of component cell kept in 80 degrees below zero and melt it in ice. (We used the compotent cell of One Shot Top 10 Chemically compotent E. coli from Invitrogen.)</p> |
| + | <p>b. Add 1㎕ of plasmid DNA to each of the tube.</p> | ||
| + | <p>c. Mix well and place them in ice for 30 minutes.</p> | ||
| + | <p>d. Place plasmid DNA in the compotent cell and keep it in 42℃ for 30 seconds and place it in ice for 2 minutes.</p> | ||
| + | <p>e. Add 250㎕ of liquid LB medium in each of the tube and grow it in 37℃ shaking incubator for an hour. </p> | ||
| + | <p>f. Leave 100㎕ and dispose.</p> | ||
| + | <p>g. Resuspend the cell, and spread 25㎕ in each of the ampicillin, kanamycin, gentamycin, tetracycline LB solid medium.</p> | ||
| + | <p>h. Wait until the E. coli bacteria smears into the plate.</p> | ||
| + | <p>i. Grow it in the 37℃ incubator for a day and observe.</p> | ||
</div> | </div> | ||
</div> | </div> | ||
Revision as of 14:25, 22 May 2014
a. Take out 50㎕of component cell kept in 80 degrees below zero and melt it in ice. (We used the compotent cell of One Shot Top 10 Chemically compotent E. coli from Invitrogen.)
b. Add 1㎕ of plasmid DNA to each of the tube.
c. Mix well and place them in ice for 30 minutes.
d. Place plasmid DNA in the compotent cell and keep it in 42℃ for 30 seconds and place it in ice for 2 minutes.
e. Add 250㎕ of liquid LB medium in each of the tube and grow it in 37℃ shaking incubator for an hour.
f. Leave 100㎕ and dispose.
g. Resuspend the cell, and spread 25㎕ in each of the ampicillin, kanamycin, gentamycin, tetracycline LB solid medium.
h. Wait until the E. coli bacteria smears into the plate.
i. Grow it in the 37℃ incubator for a day and observe.
