Team:Warsaw/Calendar-Main/23 July 2008

Cloning omega-A fusion on pKS (second attempt) Michał L., Ewa, Marcin  Isolation of plasmids from transformants. Digest of plasmids with SacI and NotI (BamHI buffer). Gel electrophoresis of products of digest. 2 selected clones were send to DNA sequencing lab. 

Cloning of protein Z DNA to <a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega>pET15b-OmpA-omega</a> in place of OmpA Paweł Ligations <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">transformed</a> into <a href="http://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> and plated on LB + ampicillin.

Preparation of chemocompetent bacteria Antoni Setup of overnight culture E. coli <a href="http://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> (LB broth) for preparation of chemocompetent bacteria.

Cloning of protein A DNA to OmpA constructs Michał K.  <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation of plasmids</a> from cultures inocluated on previous day (<a href=http://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-alpha>pACYC177+OmpA_A_alpha</a>). </li> Control <a href="http://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of isolated plasmids with BamHI and SacI (BamHI buffer).</li> Gel electrophoresis - proper clone found (<a href="http://2008.igem.org/Team:Warsaw/Calendar-Main/23_July_2008#fig1">Fig. 1.</a>).</li></ol>

<img src="http://2008.igem.org/wiki/images/f/fb/23_july.jpg" width=300/></a> Fig. 1. Control SacI/BamHI digests of pACYC177+OmpA_A_alpha

1-4. digested plasmids isolated from transformants obtained previous day 5. Marker