Team:Elan Vital South Korea/l transformation

From 2014hs.igem.org

(Difference between revisions)
 
(6 intermediate revisions not shown)
Line 10: Line 10:
                     <div id="article_c">
                     <div id="article_c">
                         <h1 class="title">Transformation</h1>
                         <h1 class="title">Transformation</h1>
-
                         <p class="listing">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 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>
-
                         <p class="listing">b. Add 1㎕ of plasmid DNA to each of the tube.</p>
+
                         <p class="listing">b. Add 1㎕ of plasmid DNA<i class="green">**</i> to each of the tube.</p>
                         <p class="listing">c. Mix well and place them in ice for 30 minutes.</p>
                         <p class="listing">c. Mix well and place them in ice for 30 minutes.</p>
                         <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>
                         <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>
-
                         <p class="listing">e. Add 250㎕ of liquid LB medium in each of the tube and grow it in 37℃ shaking incubator for an hour. </p>
+
                         <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>
                         <p class="listing">f. Leave 100㎕ and dispose.</p>
                         <p class="listing">f. Leave 100㎕ and dispose.</p>
-
                         <p class="listing">g. Resuspend the cell, and spread 25㎕ in each of the ampicillin, kanamycin, gentamycin, tetracycline LB solid medium.</p>
+
                         <p class="listing">g. Resuspend the cell, and spread 25㎕ in each of the ampicillin, kanamycin, gentamycin, tetracycline LB solid plate.</p>
-
                         <p class="listing">h. Wait until the E. coli bacteria smears into the plate.</p>
+
                         <p class="listing">h. Spread 25㎕ of competent E. coli in LB solid plate with the same antibiotics.</p>
-
                         <p class="listing">i. Grow it in the 37℃ incubator for a day and observe.</p>
+
                        <p class="listing">i. Wait until the E. coli bacteria smears into the plate.</p>
 +
                         <p class="listing">j. Grow it in the 37℃ incubator for a day and observe.</p>
 +
                        <p class="listing">k. Go through miniprep again.</p>
 +
                        <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>
 +
                        <img class="wiki_img" src="https://static.igem.org/mediawiki/2014hs/7/7b/Elan_vital_process_1.png" />
 +
                        <p class="wiki_caption">How transformation works</p>
 +
                        <img class="wiki_img" src="https://static.igem.org/mediawiki/2014hs/6/66/Elan_vital_protocol_1.png" />
 +
                        <p class="wiki_caption">Incubator</p>
                     </div>
                     </div>
                 </div>
                 </div>
             </div>
             </div>
-
            <div id="footer_c">
+
</html>
-
                <div class="inner_fix">
+
{{:Team:Elan_Vital_South_Korea/Template:footer}}
-
                    <p>&copy;2014 Elan Vital</p>
+
<html>  
-
                </div>
+
-
            </div>
+
     </body>
     </body>
</html>
</html>

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

Retrieved from "http://2014hs.igem.org/Team:Elan_Vital_South_Korea/l_transformation"