Team:Warsaw/Calendar-Main/22 May 2008

From 2008.igem.org

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<h3>Rifampicin Test</h3>
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<h3>Rifampicin test #2</h3>
<h4>Piotr</h4>
<h4>Piotr</h4>
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<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>
<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>Piotr</h4>
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<h4>Piotr, Michał K.</h4>
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<ol><li>
Design of primers for sequencing something we want from pMPM-T5.<br>
Design of primers for sequencing something we want from pMPM-T5.<br>
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<h4>Michał K.</h4>
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPM-T5 with transcriptional fusion</a>.</li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPM-T5 with transcriptional fusion</a>.</li>

Latest revision as of 14:07, 26 October 2008

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Rifampicin test #2

Piotr

Overnight inoculation and induction:
Top10
and Top10 carrying:
pMPM-T5-AID + 0.01% L-Ara
pTrc99a-AID + 0.5 mM IPTG
pMPM-T5-AID
pTrc99a-AID

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

Piotr, Michał K.

  1. Design of primers for sequencing something we want from pMPM-T5.
    Primers:

  2. Isolation of hypothetical pMPM-T5 with transcriptional fusion.
  3. Isolation of hypothetical pMPM-T5 with translational fusion
  4. Control digest of pMPM-T5 with transcriptional fusion with EcoRI (EcoRI buffer).
  5. Control digest of pMPM-T5 with translational fusion with BamHI (BamHI buffer).
  6. Gel electrophoresis (no proper colonies found). Fig. 1.
    7. Fig. 1. 1-DNA ladder;
    2 and 3-hypothetical pMPM-T5 with transcriptional fusion digested with EcoRI (2 clones);
    4-pMPM-T5-AID digested with EcoRI;
    5-hypothetical pMPM with translational fusion digested with BamHI;
    6-pMPM-T5-AID digested with BamHI.

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