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Team:Warsaw/Calendar-Main/6 October 2008
From 2008.igem.org
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<li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> with <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaL_N">OmpaL_N</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaP_link">OmpaP_link</a> | <li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> with <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaL_N">OmpaL_N</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaP_link">OmpaP_link</a> | ||
| - | primers on colonies from plates with transformations | + | primers on colonies from plates with transformations <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+ OmpA_omega (annealing temperature - 55°C,90 s of elongation step). No visible bands for proper product.</li> |
<li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> with <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinLSXNE">LinLSXNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlfaPSpe">AlfaPSpe</a> | <li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> with <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinLSXNE">LinLSXNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlfaPSpe">AlfaPSpe</a> | ||
| - | primers on colonies from plates with transformations | + | primers on colonies from plates with transformations <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+ link_alpha (annealing temperature - 55°C,60 s of elongation step). No visible bands for proper product.</li> |
| - | <li>Inoculation of some colonies which grown on plates with three separate transformations: | + | <li>Inoculation of some colonies which grown on plates with three separate transformations: <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha, <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega and pACYC177 + OmpA_omega_ to liquid LB + kanamycin. </li> |
| - | <li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of | + | <li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> plasmid with XbaI and PstI (Tango buffer). <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">Dephosphorylation</a> (CIAP) of plasmid.</li> |
<li> Gel electrophoresis 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 band - 2200 bp.</li> | <li> Gel electrophoresis 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 band - 2200 bp.</li> | ||
| - | <li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: | + | <li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + pLac_OmpA_omega (without EcoRI site) and self ligation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b+OmpA_omega</a> without XbaI.</li> |
</ol> | </ol> | ||
Revision as of 00:51, 26 October 2008
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Preparation of BioBricksMichał K.
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