Team:NU Kazakhstan/Notebook

From 2014.igem.org

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<p>May 12</p>
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          Titel</p>
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<p>Congratulations with the First Working Day!</p>
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<ul>May 13:
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      <p>text
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<li>Camelid's antibody p53 sequence was found</li>
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</p>
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<li>Exploring camelid antibodies:</li>
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<br>
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<p>1) Nanobodies: Natural Single-Domain Antibodies by Serge Muyldermans, March 2013</p>
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<p>2) Nanobody-based products as research and diagnostic tools by Thomas De Meyer et al, 2013</p></ul>
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<p>May 17: We recieved the reply from Dr. Fernandez lab who are ready to send the plasmid containing the zipper and HlyA sequence. Special thanks to Dr. Shipley who wrote the letter.</p>  
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<p>May 19: We had discussed the three topics, the nanobody (nb) project's protocol is ready, antimicrobial peptides (amp) and phi29 are under the process. We distributed the people from nb group to support othre projects.
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The new papers and nb presentation were uploaded on the google drive</p>
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<p>June 4: We did DNA miniprep of E. coli cultures in LB medium, which was obtained from plates with transformed plates. Cells were transformed with the parts from the kit: BBa_K608002 and BBa_E1010. The tubes with miniprep products are at +4C
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<br> <br><br><br><br><br>
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<ul>Concentrations of DNA measured with Nanodrop:
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          <p>text
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<li>Promoter: 221.3 ng/ul</li>
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<li>RFP: 348.2 ng/ul</li>
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<li>RFP from Arabinose plates: 230.2 ng/ul</li>
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</ul>
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<p>June 13:</p>
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<p>We did miniprep of DNA from sigle colony isolation (see June 11) and checked our DNA concentration on nanodrop:
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[BBa_I15016]=206.7ng/ul and [RFP control]=375.1ng/ul</p>
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<p>June 16. Dear team! I am glad to inform you that we have received stab of E. coli with the plasmid containing the zipper and HlyA sequence, and a stab of E. coli with pVDL9.3 encoding hlyB and hlyD!!!
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A big thank to Dr. Fernandez Herrero and to Dr. Shipley!</p>
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<p>June 19: We did pcr of Chloramphinecol plasmid backbones, and we used the following reagent mixture set up for 20ul:
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<li>Water-5ul</li>
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<li>High fidelity master mix-10ul</li>
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<li>Primer forward-1ul (we took from 50ul of 10uM stock)</li>
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<li>Primer reverse-1ul (we took from 50ul of 10uM stock)</li>
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<li>DNA template (0.2ng/ul)-3ul.</li>
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<li>PCR temperature set-up:</li>
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<li>Initial denaturation: 98C, 30s</li>
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<li>Denaturation: 98C, 10s</li>
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<li>Annealing: Temperature gradient from 57C to 71C, 30s</li>
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<li>Extension: 72C, 41s</li>
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<li>Final extension: 72C, 5min</li></p>
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<p>See agarose gel picture for the results.</p>
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           <p style="color:#E7E7E7"> <a href="https://2014.igem.org/wiki/index.php?
           <p style="color:#E7E7E7"> <a href="https://2014.igem.org/wiki/index.php?

Revision as of 07:53, 13 August 2014






May 12

Congratulations with the First Working Day!


    May 13:
  • Camelid's antibody p53 sequence was found
  • Exploring camelid antibodies:

    • 1) Nanobodies: Natural Single-Domain Antibodies by Serge Muyldermans, March 2013

    2) Nanobody-based products as research and diagnostic tools by Thomas De Meyer et al, 2013

May 17: We recieved the reply from Dr. Fernandez lab who are ready to send the plasmid containing the zipper and HlyA sequence. Special thanks to Dr. Shipley who wrote the letter.

May 19: We had discussed the three topics, the nanobody (nb) project's protocol is ready, antimicrobial peptides (amp) and phi29 are under the process. We distributed the people from nb group to support othre projects. The new papers and nb presentation were uploaded on the google drive

June 4: We did DNA miniprep of E. coli cultures in LB medium, which was obtained from plates with transformed plates. Cells were transformed with the parts from the kit: BBa_K608002 and BBa_E1010. The tubes with miniprep products are at +4C

    Concentrations of DNA measured with Nanodrop:
  • Promoter: 221.3 ng/ul
  • RFP: 348.2 ng/ul
  • RFP from Arabinose plates: 230.2 ng/ul

June 13:

We did miniprep of DNA from sigle colony isolation (see June 11) and checked our DNA concentration on nanodrop: [BBa_I15016]=206.7ng/ul and [RFP control]=375.1ng/ul

June 16. Dear team! I am glad to inform you that we have received stab of E. coli with the plasmid containing the zipper and HlyA sequence, and a stab of E. coli with pVDL9.3 encoding hlyB and hlyD!!! A big thank to Dr. Fernandez Herrero and to Dr. Shipley!

June 19: We did pcr of Chloramphinecol plasmid backbones, and we used the following reagent mixture set up for 20ul:

  • Water-5ul
  • High fidelity master mix-10ul
  • Primer forward-1ul (we took from 50ul of 10uM stock)
  • Primer reverse-1ul (we took from 50ul of 10uM stock)
  • DNA template (0.2ng/ul)-3ul.
  • PCR temperature set-up:
  • Initial denaturation: 98C, 30s
  • Denaturation: 98C, 10s
  • Annealing: Temperature gradient from 57C to 71C, 30s
  • Extension: 72C, 41s
  • Final extension: 72C, 5min

    • See agarose gel picture for the results.

    !!!

    Retrieved from "http://2014.igem.org/Team:NU_Kazakhstan/Notebook"