Team:Paris Saclay/Labwork

From 2014.igem.org

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* Digestion to have the insert [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_digestion here] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Digestion_of_PS_topo_plasmids here]
* Digestion to have the insert [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_digestion here] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Digestion_of_PS_topo_plasmids here]
* Ligation of the insert in the differents plasmids [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_ligation here] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Ligation Here]
* Ligation of the insert in the differents plasmids [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_ligation here] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Ligation Here]
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* Transformation [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/15#Transformation_with_the_ligation_.28futur_pPS2.2F4.29 Here] and  
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* Transformation [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/15#Transformation_with_the_ligation_.28futur_pPS2.2F4.29 Here] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/17#transformation here]
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* PCR on colony [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#PCR_Colony here]
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* PCR on colony [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#PCR_Colony here] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/18 here]
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* Checking of the insert sens [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/19#PCR_to_check_the_sens_of_the_insert here] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/23 here] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/24 here]
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* Final stock and extraction of pPS3 and pPS4 [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/25 here]
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====pPS5====
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*Preparation of the pPS4 plasmid with SalI digestion [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion here]
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*Ligation [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion here]
====PSBIC3====
====PSBIC3====

Revision as of 09:56, 14 October 2014

Contents

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 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

C - Salicylate Inducible Suppressing System

D - The Lemon Shaping

Countdown

This page is under Fabio's responsibility

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

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