Team:Warsaw/Calendar-Main/3 June 2008

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Preparation of pMPMT5-AID+AIDT7 construct

Michał K.:

  1. DNA (PCR products from previous day) gel electrophoresis and isolation of DNA from proper band.
  2. Digest of pMPM-T5 + transcription fusion with EcoRHI and HindIII.
  3. Use of T4 polymerase for blunting.
  4. DNA gel electrophoresis.
  5. Isolation of DNA from proper band.
  6. Ligation of product p.11 (pMPM-T5 + T7 RNA-polymerase).
  7. Transformation of E.coli TOP10 with ligation product.
  8. Plating transformants on LB + tetracycline.

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