Team:Warsaw/Calendar-Main/25 June 2008

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<ol>
<ol>
<li>We've run gradient <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> to obtain Alpha-link and link-A.
<li>We've run gradient <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> to obtain Alpha-link and link-A.
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<ul><li>Taq polymerase</li><li>buffer suplemented with (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub></li><li>MgCl<sub>2</sub> concentration gradient (from 1 to 4 mM)</li><li>annealing temp 55&deg;C.</li><li>elongation time 1 - 2 minutes.</li></ul></li>
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<ul><li>Taq polymerase</li><li>buffer suplemented with (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub></li><li>MgCl<sub>2</sub> concentration gradient (from 1 to 4 mM)</li><li>annealing temp 55&deg;C</li><li>elongation time 1 - 2 minutes</li></ul></li>
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<li>We have successfully amplified Alpha-link and link-A. Now it's time for PCL (Polymerase Chain Ligation) to create fusions  Alpha-A and Omega-A (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/25_June_2008#fig1">Fig. 1</a>).</li></ol>
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<li>We have successfully amplified Alpha-link and link-A (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/25_June_2008#fig1">Fig. 1</a>). Now it's time for PCL (Polymerase Chain Ligation) to create fusions  Alpha-A and Omega-A.</li></ol>
<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/8/89/PCR_gradient_Alinker_WAW.jpg" width=400/></a>
<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/8/89/PCR_gradient_Alinker_WAW.jpg" width=400/></a>

Revision as of 20:05, 20 October 2008

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Change of the reporter from pZC with B-galactosidaze to GFP or RFP

Piotr, Weronika

  1. Inoculation of 10ml LB with E.coli TOP10 carrying pZC
  2. Isolation and chemotransformation with plasmids nazwa (Gfp genetrator), nazwa (RFP generator)
  3. Plating of chemotransformants on LB+ampicillin

Preparation of constructs with OmpA protein fusions

Michał K.

Repetition of PCRs and gel-out purifications from 16 June.

PCR of Alpha-link and link-A

Michał L. Marcin and Ewa

  1. We've run gradient PCR to obtain Alpha-link and link-A.
    • Taq polymerase
    • buffer suplemented with (NH4)2SO4
    • MgCl2 concentration gradient (from 1 to 4 mM)
    • annealing temp 55°C
    • elongation time 1 - 2 minutes
  2. We have successfully amplified Alpha-link and link-A (Fig. 1). Now it's time for PCL (Polymerase Chain Ligation) to create fusions Alpha-A and Omega-A.
Fig. 1. PCR product - link-A.

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