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Team:Warsaw/Calendar-Main/8 July 2009
From 2009.igem.org
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| + | <a name="Isolation and digest of pKS-CRO (ligated and transformed into E. coli the previous day)"></a><h3> <span class="mw-headline">Digest of plasmids pLG612-1 and pVDL9 (containing components of hemolisin secretion system)</span></h3> | ||
| + | <p><br /> | ||
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| + | </p> | ||
| + | <a name="Kuba"></a><h4> <span class="mw-headline">Kuba</span></h4> | ||
| + | <ul> | ||
| + | <br /> | ||
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| + | <li><b>pKS-CRO cut with XbaI and EcoRI</b></li> | ||
| + | </ul> | ||
| + | <p><img src="https://static.igem.org/mediawiki/2009/4/4d/PKS-cro(XbaI%2C_EcoRI).JPG" /> | ||
| + | </p> | ||
| + | <ul> | ||
| + | <li>Gel (from the left)</li> | ||
| + | </ul> | ||
| + | <ol> | ||
| + | <li>GeneRuler DNA Ladder Mix #SM0333 (Fermentas)</li> | ||
| + | <li>uncut plasmid (isolate no.1)</li> | ||
| + | <li>cut plasmid (isolate no.1)</li> | ||
| + | <li>uncut plasmid (isolate no.2)</li> | ||
| + | <li>cut plasmid (isolate no.2)</li> | ||
| + | <li>uncut plasmid (isolate no.3)</li> | ||
| + | <li>cut plasmid (isolate no.3)</li> | ||
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| + | </ol> | ||
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Revision as of 21:08, 8 July 2009
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Gradient PCR Pho
Kama
Kama
Tasks:
- Amplification of phoP/phoQ
Methods:
PCR mixture's composition:
1ul pfu buffer (Fermentas), 1ul MgSO4 (Fermentas), 0,5ul primers, 0,5ul dNTPs (10 mM), 0,25ul pfu turbo polymerase, 0,5ul template DNA from Listeria, optionally: 0,75ul DMSO, solution was topped up with H2O to 10ul.
- PCR programs:
pho
4min 95°C
(30s 95°C, 1min 45-55°C, 4min 72°C)x3
(30s 95°C, 1min 55-60°C, 4min 72°C)x28
10min 72°C
~ 7°C
- Electrophoretic separation on 1% agarose gel
Results:
- Gel (from left)
- control -
- 2-6 - samples (annealing temperature increases to from the left to the right)
- 7-11 - samples with DMSO (annealing temperature increases to from the left to the right)
- M - GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
Notes:
Digest of plasmids pLG612-1 and pVDL9 (containing components of hemolisin secretion system)
Kuba
- pKS-CRO cut with XbaI and EcoRI
.JPG)
- Gel (from the left)
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
- uncut plasmid (isolate no.1)
- cut plasmid (isolate no.1)
- uncut plasmid (isolate no.2)
- cut plasmid (isolate no.2)
- uncut plasmid (isolate no.3)
- cut plasmid (isolate no.3)
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