Preparation of pMPMT5-AID+AIDT7 construct

Michał K.:

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

Preparation of pMPMT5+T7 construct

Piotr:

1. Digest of pMPM-T5 + transcription fusion with EcoRHI and HindIII (2x Tango buffer).
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.