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Team:Warsaw/Calendar-Main/22 April 2009
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<ul><li><p>PCR mixture's composition:</p> 2,5ul pfu buffer (Fermentas), 2,5ul MgSO4 (Fermentas), 1,5ul primers, 1,5ul dNTPs (10 mM), 01,ul pfu turbo polymerase (Fermentas), 1ul template DNA from Listeria, solution was topped up with H2O to 25ul. | <ul><li><p>PCR mixture's composition:</p> 2,5ul pfu buffer (Fermentas), 2,5ul MgSO4 (Fermentas), 1,5ul primers, 1,5ul dNTPs (10 mM), 01,ul pfu turbo polymerase (Fermentas), 1ul template DNA from Listeria, solution was topped up with H2O to 25ul. | ||
<p>1 repeat of every sample was made (2 different programs).</p> | <p>1 repeat of every sample was made (2 different programs).</p> | ||
| + | <p>Additionally for each sample was made version with 10X dilution of template.</p> | ||
</li> | </li> | ||
</ul> | </ul> | ||
<ul> | <ul> | ||
<li>PCR programs:</li> | <li>PCR programs:</li> | ||
| - | |||
<p>hly</p><pre align="left">90s 95°C <br/> (30s 95°C, 35s 42°C, 150s 72°C)x2 <br/> (30s 95°C, 35s 47°C, 150s 72°C)x28 <br/> 600s 72°C <br/> ~ 10°C</pre> | <p>hly</p><pre align="left">90s 95°C <br/> (30s 95°C, 35s 42°C, 150s 72°C)x2 <br/> (30s 95°C, 35s 47°C, 150s 72°C)x28 <br/> 600s 72°C <br/> ~ 10°C</pre> | ||
| + | <p>random program</p> | ||
</ul> | </ul> | ||
<ul> | <ul> | ||
Revision as of 00:59, 10 June 2009
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PCR mgtc, hly
Kamila
Tasks:
- Amplification of hly
Methods:
PCR mixture's composition:
2,5ul pfu buffer (Fermentas), 2,5ul MgSO4 (Fermentas), 1,5ul primers, 1,5ul dNTPs (10 mM), 01,ul pfu turbo polymerase (Fermentas), 1ul template DNA from Listeria, solution was topped up with H2O to 25ul.1 repeat of every sample was made (2 different programs).
Additionally for each sample was made version with 10X dilution of template.
- PCR programs:
hly
90s 95°C
(30s 95°C, 35s 42°C, 150s 72°C)x2
(30s 95°C, 35s 47°C, 150s 72°C)x28
600s 72°C
~ 10°C
random program
- Electrophoretic separation on 1% agarose gel
Results:
- Gel (from left)
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- mgtc 1 repeat
- mgtc 2 repeat
- mgtc sample -
- hly 1 repeat
- hly 2 repeat
- hly sample -
Notes:
- Inappropriate template DNA was used for mgtc (ganomic DNA from Yersinia instead of genomic DNA from Listeria).
