Michał K.:

1. Digestation of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI (2x Tango buffer).

2. Digestation of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI

3. Clean-up of digested products from p.1 and p.2

4. Digestation of pMPM-T5+AID with HindIII and SalI (2x Tango buffer).

5. Digestation of pMPM-T5 with NcoI and XbaI

6. Gel-electrophresis and gel-out of proper bands.

7. Ligation of purified products from p.1 and p.4

8. Electroporation of E.coli TOP10 with ligation product

9. Ligation of digested products from p.4 and p.5

10. Electroporation of E.coli TOP10 with ligation product

11. Transformants plating on LB + tetracycline

http://openwetware.org/images/thumb/6/61/UW_A%2BT.jpg/350px-UW_A%2BT.jpg http://openwetware.org/images/thumb/c/c7/UW_AT.jpg/350px-UW_AT.jpg