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Team:Warsaw/Calendar-Main/5 July 2009
From 2009.igem.org
(Difference between revisions)
| Line 6: | Line 6: | ||
<p>Tasks:</P> | <p>Tasks:</P> | ||
<ul> | <ul> | ||
| - | <li>Transformation of chemocompetent E. coli strain | + | <li>Transformation of chemocompetent E. coli strain DH5α</li><br/> |
<b>Comment:</b><br/> | <b>Comment:</b><br/> | ||
<p>When we found have appropriate bacteria I transformed it using the ligation reaction prepared in 04.07.09 to create pKS plasmid with p53 coding sequence</p> | <p>When we found have appropriate bacteria I transformed it using the ligation reaction prepared in 04.07.09 to create pKS plasmid with p53 coding sequence</p> | ||
</ul> | </ul> | ||
<br/> | <br/> | ||
| - | <p>Methods:</p> | + | <p>Methods:</p><ul> |
<li>thaw bacteria on the ice - 10 minuts</li> | <li>thaw bacteria on the ice - 10 minuts</li> | ||
<li>add 4 ul of ligation mixture to the bacteria</li> | <li>add 4 ul of ligation mixture to the bacteria</li> | ||
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<li>add 800 ul of SOB medium to the bacteria</li> | <li>add 800 ul of SOB medium to the bacteria</li> | ||
<li>incubation in 37°C - 1 h</li> | <li>incubation in 37°C - 1 h</li> | ||
| - | <li>Plating on selective LB medium supplemented with ampicillin, X-Gal and IPTG</li> | + | <li>Plating on selective LB medium supplemented with ampicillin, X-Gal and IPTG</li></ul> |
</html> | </html> | ||
Latest revision as of 11:56, 18 September 2009
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Cloning of p53 coding sequence
Marcin
Tasks:
- Transformation of chemocompetent E. coli strain DH5α
Comment:
When we found have appropriate bacteria I transformed it using the ligation reaction prepared in 04.07.09 to create pKS plasmid with p53 coding sequence
Methods:
- thaw bacteria on the ice - 10 minuts
- add 4 ul of ligation mixture to the bacteria
- incubation on the ice - 20 minutes
- heat shock - 1 minut, 42°C
- incubation on the ice - 2 minuts
- add 800 ul of SOB medium to the bacteria
- incubation in 37°C - 1 h
- Plating on selective LB medium supplemented with ampicillin, X-Gal and IPTG
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