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Team:EPF Lausanne/Notebook/week1
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Latest revision as of 19:47, 24 October 2010
Contents |
Week 1
12-07-2010
- OD measurement of Asaia’s culture
- Chemical competence for E.Coli DB3.1 (step : Day 3)
- PCR (Asaia ORI + primers)=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
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
14-07-2010
- Run a gel for the PCR of the previous day -> worked
- Purification of PCR’s product -> ok
- Preparation of the BBa_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
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
- Electrophoresis of resistance fragments (Works!!!) and extraction
- Purification of extracted DNA from agarose gel
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
- Transformation of E-Coli with the ligation products and plating the transformed bacteria
