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Team:Warsaw/Calendar-Main/23 May 2008
From 2008.igem.org
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<p>Michał K.:</p> | <p>Michał K.:</p> | ||
| - | <p>1. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest"> | + | <p>1. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of PCR product for transcription fusion (T7 RNA-polymerase) 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"> | + | <p>2. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</a> of PCR product for translation fusion (AID+T7 RNA-polymerase) 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"> | + | <p>4. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</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"> | + | <p>5. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">Digest</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.4 - 4850 bp, p.5 - 4250 bp).</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.4 - 4850 bp, p.5 - 4250 bp).</p> | ||
Revision as of 17:42, 1 October 2008
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Michał K.: 1. Digest of PCR product for transcription fusion (T7 RNA-polymerase) with HindIII and SalI (2x Tango buffer). 2. Digest of PCR product for translation fusion (AID+T7 RNA-polymerase) with NcoI and XbaI (1x Tango buffer). 3. Clean-up of digested products from p.1 and p.2. 4. Digest of pMPM-T5+AID with HindIII and SalI (2x Tango buffer). 5. Digest of pMPM-T5 with NcoI and XbaI (1x Tango buffer). 6. Gel-electrophresis and gel-out of proper bands (p.4 - 4850 bp, p.5 - 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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