IGEM:UW/2008/Notebook/iGEM UW Team/2008/06/03

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1. Gel electrophoresis (PCR products) and isolation from proper bands

2. Digestion with HindIII and SalI

3. Digestion of pMPM-T5 + AID with HindIII and SalI

4. Clean-up products of p.3 and p.4 reactions

5. Ligation of p.5 products

6. Transformation of E.coli TOP10 with ligation product

7. Transformants onto LB+tetracycline

8. Digestion of pMPM-T5 + transcription fusion with EcoRHI and HindIII

9. Use of T4 polimerase for blunting

10. Gel electrophoresis

11. Isolation from proper band

12. Ligation of product p.12

13. Transformation of E.coli TOP10 with ligation product

14. Transformants onto LB + tetracycline





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