Michał K.:

1. Digest of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI

2. Digest 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. Digest of pMPM-T5+AID with HindIII and SalI

5. Digest 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