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Team:Warsaw/Calendar-Main/23 May 2008
From 2008.igem.org
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| - | 1. Digest of | + | 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 | 2. Digest of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI | ||
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3. Clean-up of digested products from p.1 and p.2 | 3. Clean-up of digested products from p.1 and p.2 | ||
| - | 4. | + | 4. Digest of pMPM-T5+AID with HindIII and SalI |
| - | 5. | + | 5. Digest of pMPM-T5 with NcoI and XbaI |
| - | 6. | + | 6. Gel-electrophresis and gel-out of proper bands. |
| - | 7. | + | 7. Ligation of purified products from p.1 and p.4 |
| - | 8. | + | 8. Transformation of E.coli TOP10 with ligation product |
| - | 9. Ligation of digested products from p. | + | 9. Ligation of digested products from p.4 and p.5 |
10. Transformation of E.coli TOP10 with ligation product | 10. Transformation of E.coli TOP10 with ligation product | ||
Revision as of 20:15, 22 September 2008
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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. Transformation of E.coli TOP10 with ligation product 9. Ligation of digested products from p.4 and p.5 10. Transformation of E.coli TOP10 with ligation product 11. Transformants plating on LB + tetracycline
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