Preparation of pMPMT5-AID+AIDT7 construct

Michał K.:

1. DNA (PCR products from previous day) gel electrophoresis and isolation of DNA from proper band.

Piotr:

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