Team:GO Paris-Saclay/Notebook
Human Practices |
Lab book |
February 28th :- Participation of the team in Aperiscience March 12th :- Meeting with the German team of Marburg June 5th :- Choice of gene entanglement as project’s subject July 1st :- Interview Harris WANG, Thomasz Blazejewski and Hsing-I Ho (authors of Cameos, the software used to entangle genes) July ? th :Participation at the conception, realisation and shot of the video iGEM France + Switzerland Done by Clémentine, Inès, Alice, Delphine and Mathieu July or August ? th :
Participation at a Zoom with Santa Cruz Team Done by Clémentine, Inès, Nathan and Mathieu September 9th :- Parisian Meetup September 16th :- Interview Mathieu Falques Done by Delphine, Baptiste and Florent - Video collab France September 17th :- Interview Tristan Rossignol, Researcher at INRAE Done by Delphine and Alice September 18th :- Interview Anne Lopes Done by Mathieu, Godefroy and Florent September 24th and October 15th :- Interview Gaëlle Lelandais Done by Delphine and Alice October 3rd-4th :- Stand presentation at “La Cité des Sciences et de l’Industrie” - Service Médiation Scientifique (= City of Sciences and the Industry - Department of Scientific Mediation) in Paris. Done by Delphine, Alice, Mathieu, Godefroy, Clémentine, Baptiste and Florent |
July 6th to 10th :- Evaluation of the output sequences generated by CAMEOS and comparison to the input genes through BLAST. - Design of entangled gene sequences on benchling of our sequences ( entangled genes ) generated with CAMEOS ( add of RBS sequences ) and design of primers compatible with Golden Gate cloning (add BsaI Sites) - Send designed sequences (primers and genes) for synthesis. Done by Clémentine and Florent
July 13th to 17th :-Amplification, miniprep, and evaluation of recipient and template plasmids pBAD-MoClo, pZE12, pZA21, and pZA31. Done by Clémentine, Ines and Mathieu Stable YFPJuly 22th :- Dilution and evaluation of synthetic DNA from IDT of genes stable_YFP and ilvA_ccdB - GG reaction to clone biobricks stable_YFP or ilvA_ccdB into pBAD-MoClo (GG#1) - Transformation of E. coli DH5alpha cells with GG#1 reaction products: stable_YFP+pBAD-MoClo and ilvA_ccdB+pBAD-MoClo Done by Clémentine, Ines and Mathieu July 23th :- Start pre-cultures from colonies obtained after GG#1 transformation - Colony PCR from transformants GG#1 (colony_PCR#1) - Evaluation of amplicons by Agarose gel electrophoresis from colony_PCR#1 Done by Clémentine, Ines and Mathieu August 3rd :- PCR amplification and GG cloning of pZA31 and pBAD-MoClo inserts (GG#6) Done by Mathieu, Baptiste, Godefroy and Florent August 5th :- Comparison of protocols for GG cloning of knt_GFP and stable_YFP into pBAD-MoClo (GG#7) - Colony PCR of GG#7 transformants (colony_PCR#7) - Agarose gel electrophoresis from colony_PCR#7 Done by Mathieu, Baptiste, Godefroy and Florent August 6th :- Strike and store GG#7 transformants Done by Mathieu, Baptiste, Godefroy and Florent August 7th :- Culture of E. coli pBAD-knt_GFP and E. coli pBAD-YFP on selective media Done by Mathieu, Baptiste, Godefroy and Florent August 11th :- Colony PCR from cells transformed with GG#7 (pBAD-YFP), GG#8 (pBAD-ilvA_ccdB), and GG#8 (pBAD-knt_GFP) - Miniprep of pBAD-YFP from GG#7 clone 5 Done by Baptiste, Godefroy and Florent and Ibrahim August 12th :- Colony PCR for clones 1 to 5 from GG#7 (pBAD-YFP), (colony_PCR#11,5) -Agarose gel electrophoresis of colony_PCR#11,5 products - Miniprep of pBAD-YFP from GG#7 clone 5 - OD measurements from overnight cultures of E. coli pBAD-knt_GFP and E. coli pBAD-YFP - Preparation of cultures from strains E. coli pBAD-knt_GFP and E. coli pBAD-YFP for microscopique fluorescence observation and Clariostar observation - Assessing fluorescence of E. coli pBAD-knt_GFP and E. coli pBAD-YFP cells in liquid cultures - Sample preparation and sending for DNA sequencing: pBAD-YFP from GG#7 - Fluorescence microscopy observation 1 Done by Baptiste, Godefroy and Florent and Ibrahim August 13th :- Antibiotic susceptibility testing of E. coli pBAD-knt_GFP cells on spectinomycin gradient Done by Baptiste, Godefroy and Florent and Ibrahim August 14th :- Evaluation of fluorescence from E. coli pBAD-knt_GFP and E. coli pBAD-YFP cells by spectrometry (Clariostar) - Preparation of samples for Fluorescence microscopy - Fluorescent microscopy observation 2 Done by Baptiste, Godefroy and Florent and Ibrahim September 2nd :- Flow cytometry of E. coli pBAD-knt_GFP and E. coli pBAD-YFP cells - Test 1 September 3rd :- Flow cytometry of E. coli pBAD-knt_GFP and E. coli pBAD-YFP cells - Test 2 - Antibiotic susceptibility testing of E. coli pBAD-knt_GFP cells on spectinomycin gradient Done by Baptiste September 30th :- Comparison sequences YFP designed and YFP sequenced Done by Mathieu and Florent knt-GFPJuly 29th :- Dilution and evaluation of synthetic DNA from IDT of genes knt-GFP - GG#3 (Golden Gate) and transformation with its products Done by Florent, Ines and Mathieu July 30th :- Second transformation GG#3 products - PCR#5 from GG#3 and GG#4 products -Agarose gel electrophoresis of colony_PCR#5 products Done by Florent, Ines and Mathieu August 5th :- Comparison of protocols for GG cloning of knt_GFP and stable_YFP into pBAD-MoClo (GG#7) - Colony PCR of GG#7 transformants (colony_PCR#7) Done by Mathieu, Baptiste, Godefroy and Florent August 6th :- Sub-cloning transformation GG#7 Done by Mathieu, Baptiste, Godefroy and Florent August 7th :- Culture of colonies of sub-cloning GG#7 on selective middle + stocking + preparation tubes for PCR#9 Done by Mathieu, Baptiste, Godefroy and Florent August 10th :- Colony PCR #10 - Electrophoresis PCR colony #10 from sub-clones knt-GFP from GG#7 Done by Baptiste, Godefroy and Florent and Ibrahim August 11th :- Electrophoresis PCR colony #10 from sous clones knt-GFP from GG#7 ( 2 ) - Observation of knt-GFP and YFP strains with UV table - Miniprep pBAD knt_GFP sub-clone GG#7 Done by Baptiste, Godefroy and Florent and Ibrahim August 12th :- OD measurements from overnight cultures of E. coli pBAD-knt_GFP and E. coli pBAD-YFP - Culture by night of colonies knt-GFP and YFP for microscopique fluorescence observation and Clariostar observation + results observations - Preparation of cultures from strains E. coli pBAD-knt_GFP and E. coli pBAD-YFP for microscopique fluorescence observation and Clariostar observation - Assessing fluorescence of E. coli pBAD-knt_GFP and E. coli pBAD-YFP cells in liquid cultures - Sample preparation and sending for DNA sequencing: pBAD-YFP from GG#8 - Fluorescence microscopy observation 1 Done by Baptiste, Godefroy and Florent and Ibrahim August 13th :- Culture of colonies knt-GFP on selective middle - Culture by night of colonies knt-GFP and YFP for fluorescence microscopy - Culture of knt-GFP colonies one spectinomycin gradient Done by Baptiste, Godefroy and Florent and Ibrahim August 14th :- Evaluation of fluorescence from E. coli pBAD-knt_GFP and E. coli pBAD-YFP cells by spectrometry (Clariostar) - Preparation of samples for Fluorescence microscopy - Fluorescent microscopy observation 2 Done by Baptiste, Godefroy and Florent and Ibrahim September 2nd :- Flow cytometry of E. coli pBAD-knt_GFP and E. coli pBAD-YFP cells - Test 1 Done by Baptiste September 3rd :- Flow cytometry of E. coli pBAD-knt_GFP and E. coli pBAD-YFP cells - Test 2 - Culture of YFP and knt-GFP transformants on middles with different antibiotics concentration gradients Done by Baptiste September 30th :- Comparison sequences knt-GFP designed and sequenced Done by Mathieu and Florent |