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Team:Warsaw/Calendar-Main/30 June 2009
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <h3>PCR inv</h3> <h4>Kama</h4> <br /> <p>Tasks:</p> <ul> <li>Amplification of inv</li> </ul> <br /> <p>Methods:</p> <ul> <li><p>PCR...) |
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<p>Methods:</p> | <p>Methods:</p> | ||
<ul> | <ul> | ||
| - | <li><p>PCR mixture's composition:</P> 2, | + | <li><p>PCR mixture's composition:</P> |
| + | <pre>2,5μl pfu buffer (Fermentas) | ||
| + | 2,5μl MgSO4 (Fermentas) | ||
| + | 1,5μl primers | ||
| + | 1μl dNTPs (10 mM)(new) | ||
| + | 1μl template (Yersinia) | ||
| + | 0,5μl pfu turbo polymerase (KNGiE) | ||
| + | 1μl DMSO</pre> | ||
| + | <p>The solution was topped up with H2O to 25μl.</p> | ||
</li> | </li> | ||
</ul> | </ul> | ||
| Line 25: | Line 33: | ||
<br/> | <br/> | ||
<p>Results:</P> | <p>Results:</P> | ||
| - | <img src=""/> | + | <img src="https://static.igem.org/mediawiki/2009/8/8f/2009_06_30_inv_opisany.jpg"/> |
<ul> | <ul> | ||
| Line 39: | Line 47: | ||
<p>Notes:</p> | <p>Notes:</p> | ||
<ul> | <ul> | ||
| - | <li></li> | + | <li>No product</li> |
</ul> | </ul> | ||
<br /> | <br /> | ||
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| + | <html> | ||
| + | <h3>Cloning of mitochondrial targeting signal</h3> | ||
| + | <h4>Sebastian</h4> | ||
| + | <br /> | ||
| + | <p>Tasks:</p> | ||
| + | <ul> | ||
| + | <li>Transformation of E. coli DH5α</li> | ||
| + | <p>Methods:</p> | ||
| + | <p>transformation of chemocompetent DH5α E.coli cells with 100ng of pAcGFP1-Mito</p> | ||
| + | <p>Plating on LB medium supplemented with Kanamycin</p> | ||
| + | </html> | ||
Latest revision as of 18:00, 17 September 2009
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PCR inv
Kama
Tasks:
- Amplification of inv
Methods:
PCR mixture's composition:
2,5μl pfu buffer (Fermentas) 2,5μl MgSO4 (Fermentas) 1,5μl primers 1μl dNTPs (10 mM)(new) 1μl template (Yersinia) 0,5μl pfu turbo polymerase (KNGiE) 1μl DMSO
The solution was topped up with H2O to 25μl.
- PCR programs:
inv
4min 95°C
(30s 95°C, 1min 54°C , 4min15s 72°C)x31
10min 72°C
~ 7°C
- Electrophoretic separation on 1% agarose gel
- Gel (from left)
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- inv
- inv control -
- No product
- Transformation of E. coli DH5α
Results:
Notes:
Cloning of mitochondrial targeting signal
Sebastian
Tasks:
Methods:
transformation of chemocompetent DH5α E.coli cells with 100ng of pAcGFP1-Mito
Plating on LB medium supplemented with Kanamycin
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