Team:Warsaw/Calendar-Main/23 June 2008

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<h3> Blue/white and rifampicin test #2 </h3>
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<h4>Piotr</h4>
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<p>Analysis of lacZ sequence from <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a>:<br>
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There is no mutation in lacZ gene. Why are the colonies white? Reason of white phenotype unknown; this might be chromosomal mutation, transcription supression or something else). </p>
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<p>Sequencing determined that all white colonies have intact lacZ gene. Reason of white phenotype unknown. We suppose that it is due to chromosomal mutation. We need another reporter system (not splitted to chromosomal and plasmid part --> GFP? ). </p>
 
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<h3>Preparation of constructs: OmpA_alpha and OmpA_omega</h3>
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<h4>Michał K.</h4>
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Inoculation other separate transformant colonies from plates with ligations: <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-alpha>pACYC177+OmpA_alpha </a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega>pACYC177+OmpA_omega</a> (liquid LB + kanamycin).</p>
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<h3>Preparation of alpha-A and omega-A fusions</h3>
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<h4>Michał L.</h4>
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<p>We have successfully amplified Omega-linker (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/23_June_2008#fig1">Fig. 1</a>), but were unable to obtain the other. two PCRs were repeated. New annealing temp.: 48&deg;C. Mg<sup>2+</sup> concentration elevated to 4 mM.</p>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/3/36/PCR_Alinker_Omegalinker_WAW.jpg"width=200/></a>
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<var><b>Fig. 1.</b>PCR products. Lane 1 - lack of product for A-linker; lane 3 - product for Omega-linker.</var>
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Latest revision as of 15:00, 26 October 2008

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Blue/white and rifampicin test #2

Piotr

Analysis of lacZ sequence from pZC320:
There is no mutation in lacZ gene. Why are the colonies white? Reason of white phenotype unknown; this might be chromosomal mutation, transcription supression or something else).

Preparation of constructs: OmpA_alpha and OmpA_omega

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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