Team:Warsaw/Calendar-Main/5 July 2008

From 2008.igem.org

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<h3>Cloning omega-A fusion on pKS (second attempt)</h3>
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<h4>Michał L., Ewa, Marcin:</h4>
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<p>We have just received corrected version of omegaP-link10-homo2 primer. Let's hope this time everything will be fine. We are repeating the PCRs:</p><br/>
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<table id="result">
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<tr ><th colspan="4"><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a></td></tr>
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<tr><th>Product</th><th>Template</th><th>Primers</th><th>Product length</th></tr>
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<tr><th>linker-A</th><td>pDRIVE-TapTag</td><td><a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+link10+homo2">AL+link10+homo2</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI">AP+NotI</a> </td><td>470 bp</td></tr>
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<tr><th>omega-linker</th><td>pUC19</td><td><a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmegaL+SacI">OmegaL+SacI</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmegaP+link10+homo2">OmegaP+link10+homo2</a></td><td>400 bp</td></tr>
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</table>
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<h4 style="test-align: center">PCR program for linker-A and omega-linker</h4>
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<table id="result">
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<tr><th>Temperature</th><th>Time</th></tr>
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<tr><td>94&deg;C</td><td>4:00</td></tr>
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<tr><td>94&deg;C</td><td>0:30</td><td rowspan="3">28 cycles</td></tr>
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<tr><td>gradient 48-55&deg;C</td><td>0:45</td></tr>
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<tr><td>72&deg;C</td><td>0:50</td></tr>
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<tr><td>72&deg;C</td><td>10:00</td></tr>
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<tr><td>4&deg;C</td><td>infinite</td></tr>
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</table>
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Revision as of 13:48, 10 October 2008

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Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin:

We have just received corrected version of omegaP-link10-homo2 primer. Let's hope this time everything will be fine. We are repeating the PCRs:


PCR
ProductTemplatePrimersProduct length
linker-ApDRIVE-TapTagAL+link10+homo2 and AP+NotI 470 bp
omega-linkerpUC19OmegaL+SacI and OmegaP+link10+homo2400 bp

PCR program for linker-A and omega-linker

TemperatureTime
94°C4:00
94°C0:3028 cycles
gradient 48-55°C0:45
72°C0:50
72°C10:00
4°Cinfinite