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Team:Warsaw/Calendar-Main/23 May 2008
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<p>1. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI (2x Tango buffer).</p> | <p>1. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI (2x Tango buffer).</p> | ||
| - | <p>2. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI</p> | + | <p>2. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI (1x Tango buffer).</p> |
| - | <p>3. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested products from p.1 and p.2</p> | + | <p>3. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested products from p.1 and p.2.</p> |
<p>4. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of pMPM-T5+AID with HindIII and SalI (2x Tango buffer).</p> | <p>4. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of pMPM-T5+AID with HindIII and SalI (2x Tango buffer).</p> | ||
| - | <p>5. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with NcoI and XbaI</p> | + | <p>5. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digestation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with NcoI and XbaI (1x Tango buffer).</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.1 - 4850 bp, p.2 - 4250 bp).</p> |
<p>7. Ligation of purified products from p.1 and p.4</p> | <p>7. Ligation of purified products from p.1 and p.4</p> | ||
Revision as of 22:50, 30 September 2008
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Michał K.: 1. Digestation of PCR product for transcription fusion (T7 RNA-polimerase) with HindIII and SalI (2x Tango buffer). 2. Digestation of PCR product for translation fusion (AID+T7 RNA-polimerase) with NcoI and XbaI (1x Tango buffer). 3. Clean-up of digested products from p.1 and p.2. 4. Digestation of pMPM-T5+AID with HindIII and SalI (2x Tango buffer). 5. Digestation of pMPM-T5 with NcoI and XbaI (1x Tango buffer). 6. Gel-electrophresis and gel-out of proper bands (p.1 - 4850 bp, p.2 - 4250 bp). 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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