Team:Warsaw/Calendar-Main/4 June 2008

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<html><h3>Preparation of pMPMT5-AID+AIDT7 construct</h3>
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<html><h3>Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID%2BAID-T7>pMPMT5-AID+AIDT7</a> construct</h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>

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Preparation of pMPMT5-AID+AIDT7 construct

Michał K.

  1. Digest of DNA fragment (AIDT7) with HindIII and SalI (2x Tango buffer).
  2. Digest of pMPM-T5 + AID with HindIII and SalI(2x Tango buffer), dephosphorylation (CIAP).
  3. Clean-up of reaction products.
  4. Ligation of purified DNA (pMPM-T5 + AID with AID-T7 RNA-polymerase translation fusion).
  5. Transformation of E.coli TOP10 with ligation product.
  6. Plating transformants on LB+tetracycline.

Preparation of pMPMT5+T7 construct

Piotr

Inoculation some separate transformant colonies (liquid LB + tetracycline).

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