Team:EPF Lausanne/FirstWeek

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<h3>Monday 12.07.2010</h3>
<h3>Monday 12.07.2010</h3>

Revision as of 15:43, 16 July 2010

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Contents

Monday 12.07.2010

  • OD measurement of Asaia’s culture
  • Chemical competence for E.Coli DB3.1 (step : Day 3)
  • PCR (Asaia ORI + primers)
  • Made GLY agar plates without antibiotics -> failed (medium was too old)
  • Asaia O/N culture without antibiotics in order to recover some WT
  • Text for the sponsor

Tuesday 13.07.2010

  • Chemical competence for E.Coli DB3.1 (step : Day 4) -> stored at -80°C
  • Preparation of GLY Medium / LB Medium / SOC Medium / TAE 1X / TAE 0.5X
  • Run a gel for the PCR -> failed (mix of two different kits)
  • Preparation of plates (GLY Agar / LB+Kan Agar / LB+Amp Agar)
  • Learned to autoclave
  • Restarted PCR (Asaia ORI + primers)
  • Plated Asaia (O/N culture)
  • Choice of Antibiotics Biobricks resistance (Kan / Amp / Tet / Chl) from the iGem Kit and preparation of them (streak out)
  • Transformation of E.coli DB3.1 with pUC19 to check its competence
  • Transformation of E.coli DH5 with p1003 (Kan) / p1002 (Amp) / p1004 (Chl) / p1005 (Tet)
  • E.coli DB3.1 and DH5 were plated on LB+Amp Agar plates (Kan was plated on LB+Kan Agar plate)
  • Ordered material
  • Wiki brainstorming
  • Protocols printing and organization

Wednesday 14.07.2010

  • Run a gel for the PCR of the previous day -> worked
  • Purification of PCR’s product -> ok
  • Preparation of the BBα_151020 (streak out)
  • Competence Calculation of E.Coli DB3.1 (+pUC10, Amp) -> ok
  • Look at Asaia with microscope -> we have cells (pictures)
  • Growth Curve for Asaia (OD measurement each hour) -> both manually (ok) + with the plate reader
  • Plated Asaia to recover some WT (streak out)
  • Preparation of Asaia Culture (for Lemaître experiment) with Kan
  • Protocol LateX template
  • O/N culture of the E.coli DH5 transformed with the BB Antibiotics resistance (4x) -> miniprep
  • Learning HTML for the wiki
  • Text for the project presentation due to iGem
  • Writing Protocols of basic procedures for the wiki page
  • Maxi-prep of Asaia + Kan

Thursday 15.07.2010

  • MaxiPrep of Lemaître’s culture
  • Purification of the BB resistances from the E.Coli cultures
  • Protocol for cloning the Asaia Vector
  • Enzyme digestion of the BB resistances and Asaia_ORI
  • Cloning (Asaia_ORI + Vector/ Asaia_ORI + BB + Vector)
  • Wiki text
  • Preparation of LB agar + Cm
  • PCR of resistance (Works!!!) and extraction
  • Purification of extracted DNA from agarose gel

Friday 16.07.2010

  • ligation Asaia origin BB + vector
  • ligation Asaia origin BB + Kan + vector
  • ligation Asaia origin BB + Amp + vector
  • ligation Asaia origin BB + Cm + vector
  • ligation Asaia origin BB + Tet + vector
  • Extraction of the OD curve data (values)
  • Replating Asaia on plates with and without kanamicin in order to recover the WT Asaia (only WT do not grow on kanamicin)
  • Feeding and infection of Drosophila (WT and immunodeficient redish mutants) with bacteria Ecc15 and Asaia in Prof. Lemaitre Lab
  • Fluorescence microscopy -> presence of fluorescence in the body/gut as expected
Retrieved from "http://2010.igem.org/Team:EPF_Lausanne/FirstWeek"