Team:Elan Vital South Korea/l transformation

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                        <h1 class="title">Transformation</h1>
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                        <p class="listing">a. Take out 50㎕of component cell<i class="green">*</i> 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>
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                        <p class="listing">b. Add 1㎕ of plasmid DNA<i class="green">**</i> to each of the tube.</p>
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                        <p class="listing">c. Mix well and place them in ice for 30 minutes.</p>
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                        <p class="listing">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>
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                        <p class="listing">e. Add 250㎕ of liquid LB broth in each of the tube and grow it in 37℃ shaking incubator for an hour. </p>
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                        <p class="listing">f. Leave 100㎕ and dispose.</p>
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                        <p class="listing">g. Resuspend the cell, and spread 25㎕ in each of the ampicillin, kanamycin, gentamycin, tetracycline LB solid plate.</p>
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                        <p class="listing">h. Spread 25㎕ of competent E. coli in LB solid plate with the same antibiotics.</p>
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                        <p class="listing">i. Wait until the E. coli bacteria smears into the plate.</p>
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                        <p class="listing">j. Grow it in the 37℃ incubator for a day and observe.</p>
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                        <p class="listing">k. Go through miniprep again.</p>
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                        <p class="listing_sub green">*We used the competent cell from ‘One Shot® TOP10 Chemically competent E. coli’ from Introgen.</p>
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                        <p class="listing_sub green">**We used the plasmid DNA we got from miniprep previously.</p>
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                        <img class="wiki_img" src="https://static.igem.org/mediawiki/2014hs/7/7b/Elan_vital_process_1.png" />
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                        <p class="wiki_caption">How transformation works</p>
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                        <img class="wiki_img" src="https://static.igem.org/mediawiki/2014hs/6/66/Elan_vital_protocol_1.png" />
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                         <p class="wiki_caption">Incubator</p>
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Latest revision as of 20:06, 20 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 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

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