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Team:Elan Vital South Korea/l transformation
From 2014hs.igem.org
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 broth 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 plate.
h. Spread 25㎕ of competent E. coli in LB solid plate with the same antibiotics.
i. Wait until the E. coli bacteria smears into the plate.
j. Grow it in the 37℃ incubator for a day and observe.
k. 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
