Team:Warsaw/Calendar-Main/28 July 2008

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Cloning omega-A fusion on pKS (second attempt)

Michał L., Ewa, Marcin:

Sequencing confirms that we have obtained construct with omega-A fusion, but A in our constructs turned out to be trunctated and contain only one from two highly similar domains.

Checking if omp_omega_A_alpha gives ampicillin resistance
Piotr

Inoculation of omp_omega_A_alpha into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75, 1 mmol/mL

Checking the expression of omp_omega_A_alpha and omp_A_alpha
Piotr

Inoculation of omp_omega_A_alpha and omp_A_alpha with inductor (0,5 mmol/mL IPTG) and negative control without IPTG.

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 <ol><li>Inoculation of omp_omega_A_alpha and omp_A_alpha with inductor (0,5 mmol/mL IPTG) and negative control without IPTG.</li></ol>
-<br>
-<ol><li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of pACYC177+OmpA_omega_deltaA_alpha (from 25 July) with BamHI and NotI. DNA ends blunting with Klenow fragment (3 hr). </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 - 4300 bp. </li>
-<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#ligation">Ligation</a> of isolated DNA fragment (1 hr). </li>
-<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a>of E. coli <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with ligation. </li>
-<li>Transformants plating on LB + kanamycin. </li></ol>