Team:EPF Lausanne/SecondWeek

From 2010.igem.org

(Difference between revisions)
(New page: [https://2010.igem.org/Team:EPF_Lausanne home] <h3>Monday 12.07.2010</h3> *OD measurement of Asaia’s culture *Chemical competence for E.Coli DB3.1 (step : Day 3) *PCR (Asaia ORI + prime...)
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[https://2010.igem.org/Team:EPF_Lausanne home]
[https://2010.igem.org/Team:EPF_Lausanne home]
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<h3>Monday 12.07.2010</h3>
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<h3>Monday 19.07.2010</h3>
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*OD measurement of Asaia’s culture
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*Results of the transformation checked: successful!!
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*Chemical competence for E.Coli DB3.1 (step : Day 3)
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*Amplification of colonies (goal to do a miniprep of the plasmids --ori-- and --ori-resistance--)
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*PCR (Asaia ORI + primers)  
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*Results of plating of asaia culture (where we hope to recover the WT) checked: Asaia cultures grew on Kan plates also! Checked colonies for fluorescance under the microscope: no fluorescance --> Very unexpected! Don't know reason yet. 
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*Made GLY agar plates without antibiotics -> failed (medium was too old)
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*Re-grow the colonies. Multiple cycles: hope that we get WT.
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*Asaia O/N culture without antibiotics in order to recover some WT
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*Preparation of Asaia cultures for Lemaitre experiments/WT/Competence...
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*Text for the sponsor
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*Preparation of SOC medium and GLY medium
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*Autoclaving of media and flasks
<h3>Tuesday 13.07.2010 </h3>
<h3>Tuesday 13.07.2010 </h3>
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*Chemical competence for E.Coli DB3.1 (step : Day 4) -> stored at -80°C
 
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*Preparation of GLY Medium / LB Medium / SOC Medium / TAE 1X / TAE 0.5X
 
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*Run a gel for the PCR -> failed (mix of two different kits)
 
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*Preparation of plates (GLY Agar / LB+Kan Agar / LB+Amp Agar)
 
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*Learned to autoclave
 
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*Restarted PCR (Asaia ORI + primers)
 
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*Plated Asaia (O/N culture)
 
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*Choice of Antibiotics Biobricks resistance (Kan / Amp / Tet / Chl) from the iGem Kit and preparation of them (streak out)
 
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*Transformation of E.coli DB3.1 with pUC19 to check its competence
 
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*Transformation of E.coli DH5 with p1003 (Kan) / p1002 (Amp) / p1004 (Chl) / p1005 (Tet)
 
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*E.coli DB3.1 and DH5 were plated on LB+Amp Agar plates (Kan was plated on LB+Kan Agar plate)
 
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*Ordered material
 
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*Wiki brainstorming
 
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*Protocols printing and organization
 

Revision as of 14:41, 19 July 2010

home

Monday 19.07.2010

  • Results of the transformation checked: successful!!
  • Amplification of colonies (goal to do a miniprep of the plasmids --ori-- and --ori-resistance--)
  • Results of plating of asaia culture (where we hope to recover the WT) checked: Asaia cultures grew on Kan plates also! Checked colonies for fluorescance under the microscope: no fluorescance --> Very unexpected! Don't know reason yet.
  • Re-grow the colonies. Multiple cycles: hope that we get WT.
  • Preparation of Asaia cultures for Lemaitre experiments/WT/Competence...
  • Preparation of SOC medium and GLY medium
  • Autoclaving of media and flasks

Tuesday 13.07.2010

Retrieved from "http://2010.igem.org/Team:EPF_Lausanne/SecondWeek"