Team:Warsaw/Calendar-Main/10 October 2008
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<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with above ligation.</li> | <li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> with above ligation.</li> | ||
<li> Tranformants plating on LB with kanamycin. </li></ol> | <li> Tranformants plating on LB with kanamycin. </li></ol> | ||
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+ | <h3>Preparation of AraC+pBAD+AID</h3> | ||
+ | <h4>Michał K., Piotr</h4> | ||
+ | <ol> | ||
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+ | <li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmid from culture inoculated on previous day - <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> without EcoRI site.</li> | ||
+ | <li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmids with EcoRI and BamHI (BamHI buffer) - proper clones found.</li> | ||
+ | <li>Temperature gradient <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> without EcoRI site plasmid using | ||
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+ | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AraCl">AraCl</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDP_HindSpeNotPst">AIDP_HindSpeNotPst</a> | ||
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+ | primers (annealing temperature 40 - 60 °C; elongation length 2.30 min) to obtain AraC+pBAD+AID fragment. Gel electrophoresis.</li> | ||
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+ | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> without EcoRI site plasmid using | ||
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+ | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AraCl">AraCl</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDP_HindSpeNotPst">AIDP_HindSpeNotPst</a> | ||
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+ | primers (annealing temperature 45 °C; elongation length 2.30 min) to obtain AraC+pBAD+AID fragment. </li> | ||
+ | <li>Gel electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band 1800 bp. </li> | ||
+ | <li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>Digest</a> of | ||
+ | isolated PCR product with XbaI and PstI (Tango buffer). </li> | ||
+ | <li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested PCR product. </li></ol> | ||
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</ol> | </ol> | ||
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</html> | </html> |
Revision as of 11:09, 27 October 2008
Visit in US EmbassyVisas have been accorded to the whole team. Preparation of alpha_linker under PT7 (BBa_K103019)Michał K., Piotr
Preparation of pT7_omega_linkMichał K., Piotr
Preparation of AraC+pBAD+AIDMichał K., Piotr
Preparation of BioBricksMichał K.
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