Team:Paris Saclay/Labwork

From 2014.igem.org

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m (The E. coli odor-free chassis)
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===The Lemon Scent===
===The Lemon Scent===
====Preparation====
====Preparation====
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* Rehydration of BioBricks  BBa_J45014, BBa_K517003 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 here]
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* Rehydration of BioBricks  BBa_J45014, BBa_K517003 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 30th June]
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* Preparation of electrocompetent DY330 and transformation via pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/18#Preparation_of_electrocompetent_DY330_and_transformation_via_pJBEI6409 here]
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* Preparation of electrocompetent DY330 and transformation via pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/18#Preparation_of_electrocompetent_DY330_and_transformation_via_pJBEI6409 18th July]
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* Extraction of p cola plasmid DNA - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/22#Extraction_of_p_cola_plasmid_DNA here]
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* Extraction of p cola plasmid DNA - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/22#Extraction_of_p_cola_plasmid_DNA 22nd July]
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* Extraction and electrophoresis of BBa_K762100 with pSB1C3 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/24#Plasmid_DNA_Extraction_&_Electrophoresis here]
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* Extraction and electrophoresis of BBa_K762100 with pSB1C3 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/24#Plasmid_DNA_Extraction_&_Electrophoresis 24th July]
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* Gel electrophoresis of p cola - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/24#Gel_electrophoresis_of_p_cola here]
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* Gel electrophoresis of p cola - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/24#Gel_electrophoresis_of_p_cola 24th July]
====pPS1====
====pPS1====
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* PCR targeting with DY330 and pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/22#PCR_Targeting here]
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* PCR targeting with DY330 and pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/22#PCR_Targeting 22nd July]
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* Transformation of DY330 with pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/25#Transformation_of_electrocompetent_cells here]
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* Transformation of DY330 with pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/25#Transformation_of_electrocompetent_cells 25th July]
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* Liquid culture of DY330 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/28#Liquid_Culture here]
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* Liquid culture of DY330 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/28#Liquid_Culture 28th July]
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* Transformation of DY330 with pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/29#Transformation_of_electrocompetent_cells 29th here]
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* Transformation of DY330 with pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/29#Transformation_of_electrocompetent_cells 29th July]
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* Culture of DY330 + pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/31#D_-_Lemon_scent here]
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* Culture of DY330 + pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/July/31#D_-_Lemon_scent 31st July]
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* Transformation of DY330 with pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/1#New_Transformation_of_electrocompetent_cells here]
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* Transformation of DY330 with pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/1#New_Transformation_of_electrocompetent_cells 1st July]
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* Electroporation of DY330+pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/4#Electroporation here]
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* Electroporation of DY330+pJBEI-6409 - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/4#Electroporation 4th July]
====pPS2-pPS3-pPS4====
====pPS2-pPS3-pPS4====
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*PCR of the differents genes or Biobrick [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/5#PCR_LS_PS_GS_Cad Here]and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/8#Checking_PCR here]
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*PCR of the differents genes or Biobrick [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/5#PCR_LS_PS_GS_Cad 5th Septembre] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/8#Checking_PCR 8th September]
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* Cloning in Topo vector and transformation of competent E.coli [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/8#Cloning here] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#ligation_of_PS_in_TOPO_vector_and_transformation here]
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* Cloning in Topo vector and transformation of competent E.coli [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/8#Cloning 8th September] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#ligation_of_PS_in_TOPO_vector_and_transformation 11th September]
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* Checking of the cloning [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/9 here] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/12 here]
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* Checking of the cloning [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/9 9th September] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/12 12th September]
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* Plasmids extraction [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/10#Plasmid_extraction here]
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* Plasmids extraction [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/10#Plasmid_extraction 10th Septembre]
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* Digestion to have the insert [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_digestion here] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Digestion_of_PS_topo_plasmids here]
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* Digestion to have the insert [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_digestion 11th September] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Digestion_of_PS_topo_plasmids 16th September]
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* Ligation of the insert in the differents plasmids [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_ligation here] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Ligation Here]
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* Ligation of the insert in the differents plasmids [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_ligation 11th September] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Ligation 16th September]
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* Transformation [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/15#Transformation_with_the_ligation_.28futur_pPS2.2F4.29 Here] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/17#transformation here]
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* Transformation [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/15#Transformation_with_the_ligation_.28futur_pPS2.2F4.29 15th September] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/17#transformation 17th September]
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* PCR on colony [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#PCR_Colony here] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/18 here]
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* PCR on colony [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#PCR_Colony 16th September] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/18 18th September]
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* Checking of the insert sens [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/19#PCR_to_check_the_sens_of_the_insert here] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/23 here] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/24 here]
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* Checking of the insert sens [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/19#PCR_to_check_the_sens_of_the_insert 19th September] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/23 here] and [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/24 24th September]
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* Final stock and extraction of pPS3 and pPS4 [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/25 here]
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* Final stock and extraction of pPS3 and pPS4 [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/25 25 September]
====pPS5====
====pPS5====
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*Preparation of the pPS4 plasmid with SalI digestion [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion here]
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*Preparation of the pPS4 plasmid with SalI digestion [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion 24th September]
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*Ligation [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion here]
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*Ligation [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion 24 September]
====PSBIC3====
====PSBIC3====
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*Preparation of the plasmids [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#Preparation_of_pSCBIC3_plasmid here]
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*Preparation of the plasmids [http://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#Preparation_of_pSCBIC3_plasmid 11 September]
===Salicylate Inducible Suppressing System===
===Salicylate Inducible Suppressing System===
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===The Lemon Shaping===
===The Lemon Shaping===
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*Concentration Agar Test - [http://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 here]
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*Concentration Agar Test - [http://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 30th June]
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*Concentration Agar Test - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/5 here]
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*Concentration Agar Test - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/5 5th August]
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*Agar mold test - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/7 here]
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*Agar mold test - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/7 7th August]
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*Agar mold test - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/8 here]
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*Agar mold test - [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/8 8th August]
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*Incubation of E. Coli with plasmid FNR RBS AmylCP in LB [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/11 here]
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*Incubation of E. Coli with plasmid FNR RBS AmylCP in LB [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/11 11th August]
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*Coloration of agar [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/12 here]
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*Coloration of agar [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/12 12th August]
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*Coloration of agar [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/13 here]
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*Coloration of agar [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/13 13th August]
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*Preparation of M63 medium [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/14 here]
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*Preparation of M63 medium [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/14 14th August]
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* Incubation of E. Coli with plasmid FNR RBS AmylCP in M63 [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/18#Monday_18th_August here]
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* Incubation of E. Coli with plasmid FNR RBS AmylCP in M63 [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/18 18th August]
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*Transformation of odor free E. coli with plasmids coding Fluo Protein [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/27 here]
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*Transformation of odor free E. coli with plasmids coding Fluo Protein [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/27 27th August]
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* Results of Fluorescent Protein [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/29#Friday_29th_August here]
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* Results of Fluorescent Protein [http://2014.igem.org/Team:Paris_Saclay/Notebook/August/29 29th August]
{{Team:Paris_Saclay/default_footer}}
{{Team:Paris_Saclay/default_footer}}

Latest revision as of 02:07, 18 October 2014

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