IGEM:UW/2008/Notebook/iGEM UW Team/2008/06/03: Difference between revisions
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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 | |||
Revision as of 02:42, 11 June 2008
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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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