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Team:Warsaw/Calendar-Main/11 May 2009
From 2009.igem.org
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<html> | <html> | ||
| - | <h3>PCR pho, mgtc | + | <h3>PCR pho, mgtc, inv</h3> |
<h4>Kama</h4> | <h4>Kama</h4> | ||
<br /> | <br /> | ||
<p>Tasks:</p> | <p>Tasks:</p> | ||
<ul> | <ul> | ||
| - | <li>Amplification of pho, mgtc | + | <li>Amplification of pho, mgtc and inv</li> |
</ul> | </ul> | ||
<br /> | <br /> | ||
<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) | ||
| + | 1μl template | ||
| + | 0,5μl pfu turbo polymerase (KNGiE)</pre> | ||
| + | <p>The solution was topped up with H2O to 25μl.</p> | ||
</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> |
| - | <p>inv</p><pre align="left"> 4min | + | <p>inv</p><pre align="left"> 4min 95°C <br/> (30s 95°C, 1min 53°C , 4min15s 72°C)x31 <br/> 10min 72°C <br/> ~ 7°C |
</ul> | </ul> | ||
<ul> | <ul> | ||
<li>Electrophoretic separation on 1% agarose gel</li> | <li>Electrophoretic separation on 1% agarose gel</li> | ||
| + | <br/> | ||
<p>Results:</P> | <p>Results:</P> | ||
<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"/> | ||
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<ul> | <ul> | ||
<li>Gel (from left)</li> | <li>Gel (from left)</li> | ||
| - | 1. GeneRuler DNA Ladder Mix #SM0333 (Fermentas) | + | </ul> |
| - | 2. pho | + | <ol> |
| - | 3. pho control - | + | <li>1. GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> |
| - | 4. mgtc | + | <li>2. pho</li> |
| - | 5. mgtc control - | + | <li>3. pho control -</li> |
| - | 6. inv | + | <li>4. mgtc</li> |
| - | 7. inv control - | + | <li>5. mgtc control -</li> |
| - | + | <li>6. inv</li> | |
| - | Notes: | + | <li>7. inv control -</li> |
| - | No product. Check the template (Salmonella), polymerase (KNGiE) and dNTPs. | + | </ol> |
| + | </var> | ||
| + | <br /> | ||
| + | <p>Notes:</p> | ||
| + | <ul> | ||
| + | <li>No product. Check the template (Salmonella), polymerase (KNGiE) and dNTPs.</li> | ||
| + | </ul> | ||
| + | <br /> | ||
</html> | </html> | ||
Latest revision as of 17:39, 17 September 2009
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PCR pho, mgtc, inv
Kama
Tasks:
- Amplification of pho, mgtc and inv
Methods:
PCR mixture's composition:
2,5μl pfu buffer (Fermentas) 2,5μl MgSO4 (Fermentas) 1,5μl primers 1μl dNTPs (10 mM) 1μl template 0,5μl pfu turbo polymerase (KNGiE)
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
inv
4min 95°C
(30s 95°C, 1min 53°C , 4min15s 72°C)x31
10min 72°C
~ 7°C
- 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 -
- No product. Check the template (Salmonella), polymerase (KNGiE) and dNTPs.
Results:
Notes:
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