Team:Warsaw/Calendar-Main/2 October 2008

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Revision as of 18:59, 24 October 2008


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Isolation of plasmid from culture inoculated on previous day (RFP(?????)+Z).
Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.
Digest of pLac_OmpA_omega fragment with EcoRI and BcuI (BamHI buffer).
Gel elctrophoresis and gel-out of proper band - 1200 bp.
Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations RFP(?????)+OmpA (annealing temperature - 55°C,45 s of elongation step).
Gel electrophoresis.
Confirmed transformant colonies inoculated to liquid LB with ampicillin.
Overnight ligation of isolated DNA fragments: (3kb??????) + _alpha, (3kb????) + _omega and pACYC177 + OmpA_omega_ (both digested with BamHI and PstI - from 30 September).

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 <ol><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 (RFP(?????)+Z).</li>
 <li>Control <a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest>digest</a> of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.</li>
+<li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of pLac_OmpA_omega fragment with EcoRI and BcuI (BamHI buffer). </li>
 <li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 1200 bp. </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#OmpaL_N">OmpaL_N</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaP_link">OmpaP_link</a>