Team:Warsaw/Calendar-Main/11 July 2008

From 2008.igem.org

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<img src="https://static.igem.org/mediawiki/2008/9/9c/11_th_july.jpg" width=300 /><var>Fig. 1. Control BamHI and NotI digests  of plasmids, which were used in successful colony PCRs <b></b><br>  
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<img src="https://static.igem.org/mediawiki/2008/7/7e/11_th_july.jpg" width=300 /><var>Fig. 1. Colony PCR using: OmpaL_N and OmpaP_link primers<b></b><br>
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Upper<br>  
1. Marker<br>
1. Marker<br>
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2. slot 1. from yesterday's Omp_A_alpha colony PCR<br>
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2-13. colony PCR products carrying probable OmpA-alpha<br>
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2. slot 14. from yesterday's Omp_A_alpha colony PCR<br>
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Lower<br>
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2. slot 4. from yesterday's Omp_A_omega colony PCR<br>
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1. Marker<br>
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2. slot 10. from yesterday's Omp_A_omega colony PCR<br>
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2-13. colony PCR products carrying probable OmpA-alpha<br></var>

Revision as of 15:20, 25 October 2008

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Preparation of constructs with OmpA protein fusions

Michał K.

  1. Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations OmpA_alpha (annealing temperature - 55°C,45 s of elongation step).
  2. Gel electrophoresis.
  3. Confirmed transformant colonies inoculated to liquid LB with kanamycin.

Preparation of construct pKS with A protein

Michał L., Ewa, Marcin

  1. Colony PCR on colonies from plates with transformations pKS-A.
    Primers used: AL+SacI and AP+NotI
  2. Gel electrophoresis.
  3. Confirmed transformant colonies inoculated to liquid LB with ampicillin.

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

Piotr, Antoni

Isolation of plasmids from cultures inoculated on previous day.

Fig. 1. Colony PCR using: OmpaL_N and OmpaP_link primers
Upper
1. Marker
2-13. colony PCR products carrying probable OmpA-alpha
Lower
1. Marker
2-13. colony PCR products carrying probable OmpA-alpha