Team:Warsaw/Calendar-Main/23 June 2008

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<h3>Cloning of protein Z DNA to <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-alpha>pET15b-OmpA-alpha</a> in place of OmpA</h3>
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<h4>Weronika</h4>
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<a href=https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation>Isolation</a> of plasmids from cultures inoculated on previous day.
<h3>Preparation of constructs with OmpA protein fusions</h3>
<h3>Preparation of constructs with OmpA protein fusions</h3>

Revision as of 20:16, 19 October 2008

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Blue/white test

Piotr

The analysis of lacZ sequence from pZC:
There is no mutation in lacZ gene. Why the colonys are white??? Reason of white phenotype unknown (thisis must be chromosomal mutation, transcription supresion or something else) This is wery strange... We must make new reporter system...

Cloning of protein Z DNA to pET15b-OmpA-alpha in place of OmpA

Weronika

Isolation of plasmids from cultures inoculated on previous day.

Preparation of constructs with OmpA protein fusions

Michał K.

Inoculation other separate transformant colonies from plates with ligations: pACYC177+OmpA_alpha and pACYC177+OmpA_omega (liquid LB + kanamycin).

Preparation of alpha-A and omega-A fusions

Michał L.

We have successfully amplified Omega-linker (Fig. 1), but were unable to obtain the other. two PCRs were repeated. New annealing temp.: 48°C. Mg2+ concentration elevated to 4 mM.

Fig. 1.PCR products. Lane 1 - lack of product for A-linker; lane 3 - product for Omega-linker.

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