Team:Warsaw/Calendar-Main/11 May 2009
From 2009.igem.org
(Difference between revisions)
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<h3>PCR pho, mgtc and inv</h3> | <h3>PCR pho, mgtc and inv</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> | ||
+ | - 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 | ||
+ | </ul> | ||
+ | <ul> | ||
+ | <li>Electrophoretic separation on 1% agarose gel</li> | ||
<img src="https://static.igem.org/mediawiki/2009/b/b8/2009.05.11_-_PCR_pho%2C_mgtc_i_inv_opisany.jpg"/> | <img src="https://static.igem.org/mediawiki/2009/b/b8/2009.05.11_-_PCR_pho%2C_mgtc_i_inv_opisany.jpg"/> | ||
+ | Results: | ||
+ | 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 - | ||
+ | |||
+ | Notes: | ||
+ | No product. Check the template (Salmonella), polymerase (KNGiE) and dNTPs. | ||
</html> | </html> | ||
+ | |||
Revision as of 19:43, 25 June 2009
PCR pho, mgtc and inv
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 Results: 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 - Notes: No product. Check the template (Salmonella), polymerase (KNGiE) and dNTPs.
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