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Team:Warsaw/Calendar-Main/4 July 2009
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <h3>Cloning of p53 coding sequence</h3> <h4>Marcin</h4> <br/> <p>Tasks:</P> <ul> <li>Transformation of chemocompetent E. coli strain D...) |
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<p>We was not sure whether the chosen bacteria had been working and we transformed the bacteria with pKS plasmid</p> | <p>We was not sure whether the chosen bacteria had been working and we transformed the bacteria with pKS plasmid</p> | ||
<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 1 ul of empty pKS plasmid to the bacteria</li> | <li>add 1 ul of empty pKS plasmid 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 Ampicilin, X-Gal and IPTG</li><br/> | + | <li>Plating on selective LB medium supplemented with Ampicilin, X-Gal and IPTG</li></ul><br/> |
<b>Concluding remark:</b> | <b>Concluding remark:</b> | ||
<p>Used bacteria was wrong but now we know where are correct bacteria.</p> | <p>Used bacteria was wrong but now we know where are correct bacteria.</p> | ||
Latest revision as of 11:55, 18 September 2009
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Cloning of p53 coding sequence
Marcin
Tasks:
- Transformation of chemocompetent E. coli strain DH5alpha
We was not sure whether the chosen bacteria had been working and we transformed the bacteria with pKS plasmid
Methods:
- thaw bacteria on the ice - 10 minuts
- add 1 ul of empty pKS plasmid 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 Ampicilin, X-Gal and IPTG
Concluding remark:
Used bacteria was wrong but now we know where are correct bacteria.
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