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Team:Warsaw/Calendar-Main/12 May 2009
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| - | Lab meeting. | + | <strong>Lab meeting.</strong> |
<br/> | <br/> | ||
<h3>PCR mgtc, pho</h3> | <h3>PCR mgtc, pho</h3> | ||
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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 MgSO<sub>4</sub> (Fermentas) | ||
| + | 1,5μl primers | ||
| + | 1μl dNTPs (10 mM)(new) | ||
| + | 1μl template (Salmonella) | ||
| + | 0,5μl pfu turbo polymerase (KNGiE) | ||
| + | 1μl DMSO</pre> | ||
| + | <p>The solution was topped up with H2O to 25μl.</pre> | ||
</li> | </li> | ||
</ul> | </ul> | ||
<ul> | <ul> | ||
<li>PCR programs:</li> | <li>PCR programs:</li> | ||
| - | <p>pho</p><pre align="left"> 2min30s | + | <p>pho</p><pre align="left"> 2min30s 95°C <br/> (30s 95°C, 35s 56°C, 3min30s 72°C)x3 <br/> (30s 95°C, 35s 61°C, 3min30s 72°C)x28 <br/> 10min 72°C <br/> ~ 7°C<br/></pre> |
| - | <p>mgtc</p><pre align="left"> 1min30s | + | <p>mgtc</p><pre align="left"> 1min30s 95°C <br/> (30s 95°C, 35s 48°C, 1min 72°C)x3 <br/> (30s 95°C, 35s 58°C, 1min 72°C)x28 <br/> 10min 72°C <br/> ~ 7°C<br/></pre> |
</ul> | </ul> | ||
<ul> | <ul> | ||
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<li> mgtc</li> | <li> mgtc</li> | ||
<li> mgtc control -</li> | <li> mgtc control -</li> | ||
| - | <br/> | + | <li><br/> |
<li> GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> | <li> GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> | ||
<li> pho</li> | <li> pho</li> | ||
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<p>Notes:</p> | <p>Notes:</p> | ||
<ul> | <ul> | ||
| - | <p><b>mgtc was succesfully amplified</b></p> | + | <p><b>mgtc was succesfully amplified<font color="green"> (termocykler z gradientem w pokoju 145)</font></b></p> |
</html> | </html> | ||
Latest revision as of 17:42, 17 September 2009
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Lab meeting.
PCR mgtc, pho
Kama
Tasks:
- Amplification of pho, mgtc
Methods:
PCR mixture's composition:
2,5μl pfu buffer (Fermentas) 2,5μl MgSO4 (Fermentas) 1,5μl primers 1μl dNTPs (10 mM)(new) 1μl template (Salmonella) 0,5μl pfu turbo polymerase (KNGiE) 1μl DMSO
The solution was topped up with H2O to 25μl.
- PCR programs:
pho
2min30s 95°C
(30s 95°C, 35s 56°C, 3min30s 72°C)x3
(30s 95°C, 35s 61°C, 3min30s 72°C)x28
10min 72°C
~ 7°C
mgtc
1min30s 95°C
(30s 95°C, 35s 48°C, 1min 72°C)x3
(30s 95°C, 35s 58°C, 1min 72°C)x28
10min 72°C
~ 7°C
- Electrophoretic separation on 1% agarose gel
- Gel (from left)
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- mgtc
- mgtc control -
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- pho
- pho control -
Results:
Notes:
mgtc was succesfully amplified (termocykler z gradientem w pokoju 145)
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