Team:Warsaw/Calendar-Main/9 May 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).
  3. 1-DNA ladder; 2-pTRC99a-AID digested with NcoI and HindIII

  4. Restriction digest of pMPM-T5 with HindIII and NcoI (1x Tango buffer), dephosphorylation (CIAP).
  5. DNA electrophoresis and isolation of pMPM-T5 vector from proper band (4250 bp).
  6. 1-DNA ladder; 2-pMPM-T5 digested with NcoI and HindIII

  7. Electrophoresis to estimate the concentration of isolated DNA.
  8. Ligation of isolated DNA fragments.