Team:Warsaw/Calendar-Main/15 May 2008

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<html><h3>PCRs for fusions</h3>
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<html><h3>Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPMT5-AID+T7</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPMT5+AID-T7</a></h3>
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<h4>Michał K.:</h4>
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<h4>Michał K.</h4>
<p><ol>
<p><ol>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - AID for translational fusion. <br>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - AID for translational fusion. <br>
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Template DNA: <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99A-AID</a>
Template DNA: <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99A-AID</a>
  <br>
  <br>
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Annealing temperature: 55 °C<br>
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Annealing temperature: 55&deg;C<br>
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Elongation time: 60 sec<br>
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Elongation time: 60 s<br>
20 cycles
20 cycles
</li>
</li>
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<li> <html><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA-polymerase for translational fusion; Taq polymerase with supplemented buffer (Łukasz Banasiak helped us with that problematique PCR).<br>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA-polymerase for translational fusion; Taq polymerase with supplemented buffer (Łukasz Banasiak helped us with that problematic PCR).<br>
Primers:
Primers:
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Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a>  genomic DNA<br>
Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a>  genomic DNA<br>
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Annealing temperature: 62 °C<br>
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Annealing temperature: 62&deg;C<br>
Elongation time: 4 minutes<br>
Elongation time: 4 minutes<br>
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</li>
</li>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA-polymerase for transcription fusion.<br>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> - T7 RNA-polymerase for transcriptional fusion.<br>
Primers:
Primers:
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Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a>  genomic DNA<br>
Template DNA: <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a>  genomic DNA<br>
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Annealing temperature: 62 °C<br>
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Annealing temperature: 62&deg;C<br>
Elongation time: 4 minutes<br>
Elongation time: 4 minutes<br>

Latest revision as of 13:46, 26 October 2008

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Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Michał K.

  1. PCR - AID for translational fusion.
    Primers: AIDlNrH AIDpLinB
    Template DNA: pTrc99A-AID
    Annealing temperature: 55°C
    Elongation time: 60 s
    20 cycles
  2. PCR - T7 RNA-polymerase for translational fusion; Taq polymerase with supplemented buffer (Łukasz Banasiak helped us with that problematic PCR).
    Primers: T7lLinkB T7pXbSal
    Template DNA: E. coli Rosetta genomic DNA
    Annealing temperature: 62°C
    Elongation time: 4 minutes
    20 cycles
  3. PCR - T7 RNA-polymerase for transcriptional fusion.
    Primers: T7lRBSHi T7pXbSal
    Template DNA: E. coli Rosetta genomic DNA
    Annealing temperature: 62°C
    Elongation time: 4 minutes
    20 cycles

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