Team:Warsaw/Calendar-Main/28 May 2008

From 2008.igem.org

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<h4>Michał K.</h4>
<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 pMPM-T5 with transcriptional fusion.</li>
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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>
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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 pMPM-T5 with translational fusion.</li>
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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%2BAIDT7>pMPM-T5 with translational fusion</a>.</li>
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<li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of pMPM-T5 with transcriptional fusion with EcoRI (EcoRI buffer).</li>
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<li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPM-T5 with transcriptional fusion</a> with EcoRI (EcoRI buffer).</li>
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<li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of pMPM-T5 with translational fusion with BamHI (BamHI buffer).</li>
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<li> Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPM-T5 with translational fusion</a> with BamHI (BamHI buffer).</li>
<li> Gel electrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/28_May_2008#fig1">Fig. 1</a> and <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/28_May_2008#fig2">Fig. 2</a>). </li>
<li> Gel electrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/28_May_2008#fig1">Fig. 1</a> and <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/28_May_2008#fig2">Fig. 2</a>). </li>
<li> Choice of proper clones and preparation for sequencing.</li>
<li> Choice of proper clones and preparation for sequencing.</li>

Revision as of 15:55, 12 October 2008

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

Michał K.

  1. Isolation of hypothetical pMPM-T5 with transcriptional fusion.
  2. Isolation of hypothetical pMPM-T5 with translational fusion.
  3. Control digest of pMPM-T5 with transcriptional fusion with EcoRI (EcoRI buffer).
  4. Control digest of pMPM-T5 with translational fusion with BamHI (BamHI buffer).
  5. Gel electrophoresis (Fig. 1 and Fig. 2).
  6. Choice of proper clones and preparation for sequencing.

Rifampicin test

Michał L., Ewa, Marcin

  1. Inoculation of liquid LB with E.coli TOP10 carrying pMPM-T5+AID
  2. Inoculation of liquid LB with E.coli TOP10 carrying pTrc99A+AID


Fig. 1. Here we found the clone with translational fusion on pMPM. These are different isolations in each lane. We have been working with the AT6 ever since.
Digest was conducted with BamHI
Fig. 2. 2 to 6 - Control digests of few clones with transcriptional fusion;
7 - pMPMT5-AID; all plasmids digested with EcoRI.



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