Labwork

Planning

A compléter: We started the work in June, with the research project scope and bibliography. In early July we began the phase of experiments, which was completed in the first half of September. We students organized ourselves into four groups of summer internship. The groups were formed by virtually all members of the team, so even students from areas other than biology could cooperate with the project in the laboratory experiments. Into four working groups, the laboratory experiments. To ensure the smooth development of the project by all students, the work had a life cycle.   a cycle of "aprendisagem" >> "Training" >> 'Development' >> "Transmission of acquired knowledge."

A - The E. coli odor-free chassis

• Culture of MG1655 and MG1655Z1 strains- 30th June
• P1 phage stock preparation for the transduction of the Delta-tnaA::Kan - 2nd July
• P1 phage transduction using the stock prepared on July 2nd to MG1655 and MG1655Z1 strains - 3rd July
• Cultures of MG1655 and MG1655Z1 transductants - 4th July
• Transformations test of competent cells MG1655 and MG1655Z1 - 10th July
• Preparation of competent cells E coli, deletion of the antibiotic cassette in the odorless bacteria - 11th July
• Preparation of competent cells E. coli MG1655 and MG1655Z1 transductants - 17th July
• Transformation of supercompetent cells MG1655 and MG1655Z1 transductants with plasmid BT340- 18th July
• Results of the transformation - 21st July
• Preparation and transformation of competent MG1655 and MG1655Z1 transductants - 22nd July
• PCR verification of the strains grown - 25th July
• Preparation and Transformation of competent MG1655 and MG1655Z1 with BT340 - 28th July
• PCR of MG1655 and MG1655Z1 with FTR-Apra-F and FTR-Apra-R - 29th July
• Preparation and transformation of electrocompetent MG1655 and MG1655Z1 with plasmid BT340 - 30th July
• Clone isolation by streak of Delta-tnaA MG1655 and MG1655Z1 on LB dishes - 1st August
• Clone isolation by streak of Delta-tnaA MG1655 and MG1655Z1 on LB and kan dishes - 4th August
• PCR verification of the final strains Delta-tnaA MG1655 and MG1655Z1 - 5th August
• Final stock of MG1655 Delta-tnaA - 6th August

The Lemon Scent

Preparation

• Rehydration of BioBricks BBa_J45014, BBa_K517003 - here
• Preparation of electrocompetent DY330 and transformation via pJBEI-6409 - here
• Extraction of p cola plasmid DNA - here
• Extraction and electrophoresis of BBa_K762100 with pSB1C3 - here
• Gel electrophoresis of p cola - here

pPS1

• PCR targeting with DY330 and pJBEI-6409 - here
• Transformation of DY330 with pJBEI-6409 - here
• Liquid culture of DY330 - here
• Transformation of DY330 with pJBEI-6409 - 29th here
• Culture of DY330 + pJBEI-6409 - here
• Transformation of DY330 with pJBEI-6409 - here
• Electroporation of DY330+pJBEI-6409 - here

pPS2-pPS3-pPS4

• PCR of the differents genes or Biobrick Hereand here
• Cloning in Topo vector and transformation of competent E.coli here
• Checking of the cloning here

C - Salicylate Inducible Suppressing System

• Rehydration of BioBricks BBa_J61051 and BBa_K228001- 23rd July
• Transformation of competent E.coli cells - 24th July
• Bacterial culture - 25th July
• Plasmid DNA Purification - 28th July
• Digestion to check & Electrophoresis - 28th July
• Main Digestion of BBa_J61051 and BBa_K228001 - 29th July
• Segregate Process - 29th July
• DNA purification gel agarose - 30th July
• Ligation - 30th July
• Transformation of competent E.coli cells - 31st July
• Liquid Culture - 1st August
• Plasmid DNA Purification - 4th August
• Digestion to check & Electrophoresis - 4th August
• Final Stock BBa_K1372000 - 4th August
• Liquid Culture - 5th August
• Plasmid DNA Purification - 6th August
• Digestion of BBa_K1372000 and BBa_B0015- 6th August
• Segregate Process - 6th August
• DNA purification gel agarose - 7th August
• Ligation - 7th August
• Transformation of competent E.coli cells - 8th August
• Liquid Culture - 11th August
• Plasmid DNA Purification - 12th August
• Digestion to check & Electrophoresis - 12th August
• Final Stock BBa_K1372001 - 12th August

D - The Lemon Shaping

• Concentration Agar Test - 30th June
• Concentration Agar Test - 5th August
• Agar mold test - 7th August
• Agar mold test - 8th August
• Incubation of E. Coli with plasmid FNR RBS AmylCP in LB 11th August
• Coloration of agar 12th August
• Coloration of agar 13th August
• Preparation of M63 medium 14th August
• Incubation of E. Coli with plasmid FNR RBS AmylCP in M63 18th August
• Transformation of odor free E. coli with plasmids coding Fluo Protein 27th August
• Results of Fluorescent Protein 29th August

Countdown

This page is under Fabio's responsibility

• Deadline: 08/oct.
  • Planning of each subproject
• Deadline: 12/oct
  • Final review Solenne