Team:Warsaw/Calendar-Main/12 May 2009
From 2009.igem.org
(Difference between revisions)
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<h3>PCR mgtc, pho</h3> | <h3>PCR mgtc, pho</h3> | ||
<h4>Kama</h4> | <h4>Kama</h4> | ||
+ | <br /> | ||
+ | <p>Tasks:</p> | ||
+ | <ul> | ||
+ | <li>Amplification of pho, mgtc, inv</li> | ||
+ | </ul> | ||
+ | <br /> | ||
+ | <p>Methods:</p> | ||
+ | <ul> | ||
+ | <li><p>PCR mixture's composition:</P> 2,5ul pfu buffer (Fermentas), 2,5ul MgSO4 (Fermentas), 1,5ul primers, 1ul dNTPs (10 mM), 1ul template, 0,5ul pfu turbo polymerase (KNGiE), solution was topped up with H2O to 25ul. | ||
+ | </li> | ||
+ | </ul> | ||
+ | <ul> | ||
+ | <li>PCR programs:</li> | ||
+ | <p>pho</p><pre align="left"> 2min30s 95C <br/> 3x(30s 95C, 35s 56C, 3min30s 72C) <br/> 28x(30s 95C, 35s 61C, 3min30s 72C) <br/> 10min 72C <br/> ~ 7C<br/></pre> | ||
+ | <p>mgtc</p><pre align="left"> 1min30s 95C <br/> 3x(30s 95C, 35s 48C, 1min 72C) <br/> 28x(30s 95C, 35s 58C, 1min 72C) <br/> 10min 72C <br/> ~ 7C<br/></pre> | ||
+ | <p>inv</p><pre align="left"> 4min 95C <br/> 31x(30s 95C, 1min 53C , 4min15s 72C) <br/> 10min 72C <br/> ~ 7C | ||
+ | </ul> | ||
+ | <ul> | ||
+ | |||
+ | <li>Electrophoretic separation on 1% agarose gel</li> | ||
+ | <br/> | ||
+ | <p>Results:</P> | ||
<img src="https://static.igem.org/mediawiki/2009/2/27/2009.05.12_-_PCR_pho_i_mgtc_opisany.jpg"/> | <img src="https://static.igem.org/mediawiki/2009/2/27/2009.05.12_-_PCR_pho_i_mgtc_opisany.jpg"/> | ||
<br/> | <br/> | ||
+ | |||
+ | <ul> | ||
+ | <li>Gel (from left)</li> | ||
+ | </ul> | ||
+ | <ol> | ||
+ | <li>1. GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> | ||
+ | <li>2. pho</li> | ||
+ | <li>3. pho control -</li> | ||
+ | <li>4. mgtc</li> | ||
+ | <li>5. mgtc control -</li> | ||
+ | <li>6. inv</li> | ||
+ | <li>7. inv control -</li> | ||
+ | </ol> | ||
+ | </var> | ||
+ | <br /> | ||
+ | <p>Notes:</p> | ||
+ | <ul> | ||
<p><b>mgtc was succesfully amplified</b></p> | <p><b>mgtc was succesfully amplified</b></p> | ||
</html> | </html> |
Revision as of 20:45, 25 June 2009
Lab meeting.
PCR mgtc, pho
Kama
Tasks:
- Amplification of pho, mgtc, inv
Methods:
PCR mixture's composition:
2,5ul pfu buffer (Fermentas), 2,5ul MgSO4 (Fermentas), 1,5ul primers, 1ul dNTPs (10 mM), 1ul template, 0,5ul pfu turbo polymerase (KNGiE), solution was topped up with H2O to 25ul.
- PCR programs:
pho
2min30s 95C
3x(30s 95C, 35s 56C, 3min30s 72C)
28x(30s 95C, 35s 61C, 3min30s 72C)
10min 72C
~ 7C
mgtc
1min30s 95C
3x(30s 95C, 35s 48C, 1min 72C)
28x(30s 95C, 35s 58C, 1min 72C)
10min 72C
~ 7C
inv
4min 95C
31x(30s 95C, 1min 53C , 4min15s 72C)
10min 72C
~ 7C
- Electrophoretic separation on 1% agarose gel
- Gel (from left)
- 1. GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- 2. pho
- 3. pho control -
- 4. mgtc
- 5. mgtc control -
- 6. inv
- 7. inv control -
Results:
Notes:
mgtc was succesfully amplified
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