Team:Warsaw/Calendar-Main/5 June 2008

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<h3>Preparation of pMPMT5+T7 construct<br>
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<h3>Preparation of pMPMT5+T7 construct</h3>
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Piotr</h3><ol>  
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<h4>Piotr</h4><ol>  
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical pMPMT5 with T7 RNA polymerase.</li>
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical pMPMT5 with T7 RNA polymerase.</li>
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<h3>Preparation of pMPMT5-AID+AIDT7 construct<br>
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<h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
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Michał K.</h3>
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<h4>Michał K.</h4>
<p>Inoculation of 5 colonies of transformants (pMPMT5-AID with AID-T7 RNA-polymerase translation fusion) into liquid LB+tetracycline.</p>
<p>Inoculation of 5 colonies of transformants (pMPMT5-AID with AID-T7 RNA-polymerase translation fusion) into liquid LB+tetracycline.</p>

Revision as of 10:03, 11 October 2008

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Preparation of pMPMT5+T7 construct

Piotr

  1. Isolation of hypothetical pMPMT5 with T7 RNA polymerase.
  2. Control digest of pMPMT5 + T7 RNA-polymerase with EcoRI.
  3. Gel electrophoresis.
  4. Choice of proper clone and preparation for sequencing.
  5. Design of primers for sequencing β-galactosidase from pZC320.
    Primers: pZCseqL and pZCseqP

Preparation of pMPMT5-AID+AIDT7 construct

Michał K.

Inoculation of 5 colonies of transformants (pMPMT5-AID with AID-T7 RNA-polymerase translation fusion) into liquid LB+tetracycline.

Blue/white and rifampicin test

Michał L., Ewa, Marcin:

  1. Inoculation of of E. coli TOP10 with pZC320 into liquid LB broth (with 30 μg/ml of ampicillin).

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