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Team:Elan Vital South Korea/l transformation
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<p class="listing">i. Grow it in the 37℃ incubator for a day and observe.</p> | <p class="listing">i. Grow it in the 37℃ incubator for a day and observe.</p> | ||
<p class="listing">j. Go through miniprep again.</p> | <p class="listing">j. Go through miniprep again.</p> | ||
| - | <p class="listing_sub green">*We used the competent cell from ‘One | + | <p class="listing_sub green">*We used the competent cell from ‘One Shot® TOP10 Chemically competent E. coli’ from Introgen.</p> |
<p class="listing_sub green">**We used the plasmid DNA we got from miniprep previously.</p> | <p class="listing_sub green">**We used the plasmid DNA we got from miniprep previously.</p> | ||
<img class="wiki_img" src="https://static.igem.org/mediawiki/2014hs/7/7b/Elan_vital_process_1.png" /> | <img class="wiki_img" src="https://static.igem.org/mediawiki/2014hs/7/7b/Elan_vital_process_1.png" /> | ||
Revision as of 13:09, 17 June 2014
Transformation
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.
j. Go through miniprep again.
*We used the competent cell from ‘One Shot® TOP10 Chemically competent E. coli’ from Introgen.
**We used the plasmid DNA we got from miniprep previously.
How transformation works
Incubator
