Team:Warsaw/Calendar-Main/2 October 2008

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Preparation of BioBricks

Michał K.

Isolation of plasmids from culture inoculated on previous day (pSB1A3+Z(BBa_K103004)).
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 pSB1A3+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: pSB2K3 + _alpha, pSB2K3 + _omega and pACYC177 + OmpA_omega_ (both digested with BamHI and PstI - from 30 September).

@@ Line 7: / Line 7: @@
 <h3>Preparation of BioBricks</h3>
 <h4>Michał K.</h4>
-<ol><li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from culture inoculated on previous day (RFP(?????)+Z).</li>
+<ol><li><a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from culture inoculated on previous day (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+<a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103004>Z(BBa_K103004)</a>).</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>
@@ Line 13: / Line 13: @@
 <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 RFP(?????)+OmpA (annealing temperature - 55&deg;C,45 s of elongation step). </li>
+  primers on colonies from plates with transformations <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+OmpA (annealing temperature - 55&deg;C,45 s of elongation step). </li>
 <li> Gel electrophoresis.</li>
 <li> Confirmed transformant colonies inoculated to liquid LB with ampicillin.</li>
-<li> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: (3kb??????) + _alpha, (3kb????) + _omega and pACYC177 + OmpA_omega_ (both digested with BamHI and PstI - from 30 September).</li>
+<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> + _alpha, <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega and pACYC177 + OmpA_omega_ (both digested with BamHI and PstI - from <a href=https://2008.igem.org/Team:Warsaw/Calendar-Main/30_September_2008>30 September</a>).</li>