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."

The E. coli odor-free chassis

• Culture of MG1655 and MG1655Z1 strains- here
• P1 phage stock preparation for the transduction of the Delta-tnaA::Kan - here
• P1 phage transduction using the stock prepared on July 2nd to MG1655 and MG1655Z1 strains - here
• Cultures of MG1655 and MG1655Z1 transductants - here
• Transformations test of competent cells MG1655 and MG1655Z1 - here
• Preparation of competent cells E coli, deletion of the antibiotic cassette in the odorless bacteria - here
• Preparation of competent cells E. coli MG1655 and MG1655Z1 transductants - here
• Transformation of supercompetent cells MG1655 and MG1655Z1 transductants with plasmid BT340- here
• Results of the transformation - here
• Preparation and transformation of competent MG1655 and MG1655Z1 transductants - here
• PCR verification of the strains grown - here
• Preparation and Transformation of competent MG1655 and MG1655Z1 with BT340 - here
• PCR of MG1655 and MG1655Z1 with FTR-Apra-F and FTR-Apra-R - here
• Preparation and transformation of electrocompetent MG1655 and MG1655Z1 with plasmid BT340 - here
• Clone isolation by streak of Delta-tnaA MG1655 and MG1655Z1 on LB dishes - here
• Clone isolation by streak of Delta-tnaA MG1655 and MG1655Z1 on LB and kan dishes - here
• PCR verification of the final strains Delta-tnaA MG1655 and MG1655Z1 - here
• Final stock of MG1655 Delta-tnaA - here

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 and here
• Checking of the cloning here and here
• Plasmids extraction here
• Digestion to have the insert here and here
• Ligation of the insert in the differents plasmids here and Here
• Transformation Here and here
• PCR on colony here and here
• Checking of the insert sens here and here and here
• Final stock and extraction of pPS3 and pPS4 here

pPS5

• Preparation of the pPS4 plasmid with SalI digestion here
• Ligation here

PSBIC3

• Preparation of the plasmids here

Salicylate Inducible Suppressing System

• Rehydration of BioBricks BBa_J61051 and BBa_K228001- here
• Transformation of competent E.coli cells - here
• Bacterial culture - here
• Plasmid DNA Purification - here
• Digestion to check & Electrophoresis - here
• Main Digestion of BBa_J61051 and BBa_K228001 - here
• Segregate Process - here
• DNA purification gel agarose - here
• Ligation - 30th July
• Transformation of competent E.coli cells - here
• Liquid Culture - here
• Plasmid DNA Purification - here
• Digestion to check & Electrophoresis - here
• Final Stock BBa_K1372000 - here
• Liquid Culture - [1]
• Plasmid DNA Purification - here
• Digestion of BBa_K1372000 and BBa_B0015- here
• Segregate Process - here
• DNA purification gel agarose - here
• Ligation - 7th August
• Transformation of competent E.coli cells - here
• Liquid Culture - here
• Plasmid DNA Purification - here
• Digestion to check & Electrophoresis - here
• Final Stock BBa_K1372001 - here

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