Team:Warsaw/Calendar-Main/28 May 2008

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<html><h3>Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7<br>
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<html><h3>Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7</h3>
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Michał K.</h3>
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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 pMPM-T5 with transcriptional fusion.</li>
<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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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. Choice of proper clones and preparation for sequencing.

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
2 to 6 - Control digests of few clones with transcription fusion;
7 - pMPMT5-AID; all plasmids digested with EcoRI.

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

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