Team:Warsaw/Calendar-Main/9 May 2008

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<html><h3>Preparation of pMPMT5+AID construct</h3>
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<html><h3>Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> construct</h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>

Revision as of 16:11, 12 October 2008

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Preparation of pMPMT5+AID construct

Michał K.

  1. Restriction digest of pTrc99A-AID with HindIII and NcoI (1x Tango buffer).
  2. DNA electrophoresis and isolation of DNA encoding AID from proper band (600 bp). Fig. 1.
  3. Restriction digest of pMPM-T5 with HindIII and NcoI (1x Tango buffer), dephosphorylation (CIAP).
  4. DNA electrophoresis and isolation of pMPM-T5 vector from proper band (4250 bp). Fig. 2.
  5. Electrophoresis to estimate the concentration of isolated DNA.
  6. Ligation of isolated DNA fragments.

Fig. 1. 1-DNA ladder; 2-pTRC99a-AID digested with NcoI and HindIII Fig. 2. 1-DNA ladder; 2-pMPM-T5 digested with NcoI and HindIII






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