Team:Paris Saclay/Labwork

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Contents

Labwork

Planning

We started the labwork in June. The first step was the research project scope and bibliography to ensure the feasibility of our project and to plan all the work we had to do during this wonderful iGEM summer. In early July we began the experiments, which was completed in the end of September. In order to be effective, we organized ourselves into four groups of summer interns. The groups were formed by all the members of the team, so even students from areas other than biology could cooperate for the project in the laboratory. To ensure the smooth development of the project by all students, the work had a life cycle :

  • Learning: A non biologist had to first learn the basics of biology to understand the lab work.
  • Practice: The novice starts to do their experiments with the help of another biology student or an adviser.
  • Development: The aforementioned student becomes autonomous and takes initiative.
  • Transmission of acquired knowledge: The student, initially oblivious to practices in the lab, can transmit this knowledge at the end of their internship to newcomers.

The E. coli odor-free chassis

  • Culture of MG1655 and MG1655Z1 strains- 30st July
  • 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 - 4st 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 - 30st 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 - 6st August

The Lemon Scent

Preparation

  • Rehydration of BioBricks BBa_J45014, BBa_K517003 - 30th June
  • Preparation of electrocompetent DY330 and transformation via pJBEI-6409 - 18th July
  • Extraction of p cola plasmid DNA - 22nd July
  • Extraction and electrophoresis of BBa_K762100 with pSB1C3 - 24th July
  • Gel electrophoresis of p cola - 24th July

pPS1

  • PCR targeting with DY330 and pJBEI-6409 - 22nd July
  • Transformation of DY330 with pJBEI-6409 - 25th July
  • Liquid culture of DY330 - 28th July
  • Transformation of DY330 with pJBEI-6409 - 29th July
  • Culture of DY330 + pJBEI-6409 - 31st July
  • Transformation of DY330 with pJBEI-6409 - 1st July
  • Electroporation of DY330+pJBEI-6409 - 4th July

pPS2-pPS3-pPS4

pPS5

PSBIC3

Salicylate Inducible Suppressing System

Construction of the Fusion Color Chromoprotein

The Lemon Shaping

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