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Team:Warsaw/Calendar-Main/23 May 2008
From 2008.igem.org
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<p>4. Digest of pMPM-T5+AID with HindIII and SalI</p> | <p>4. Digest of pMPM-T5+AID with HindIII and SalI</p> | ||
| - | <p>5. Digest of pMPM-T5 with NcoI and XbaI</p> | + | <p>5. Digest of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with NcoI and XbaI</p> |
<p>6. Gel-electrophresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands.</p> | <p>6. Gel-electrophresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands.</p> | ||
Revision as of 15:36, 27 September 2008
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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
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