Team:EPF Lausanne/FirstWeek

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

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
Culture of Asaia + Kan

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
Transformation of E-Coli with the ligation products and plating the transformed bacteria

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