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Team:Paris Saclay/Labwork
From 2014.igem.org
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=Labwork= | =Labwork= | ||
==Planning== | ==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 : | |
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| - | To ensure the smooth development of the project by all students, the work had a life cycle | + | |
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| - | === | + | * Learning: A non biologist had to first learn the basics of biology to understand the lab work. |
| - | *Culture of MG1655 and MG1655Z1 strains- [https://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 | + | * 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. | ||
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| + | ===The ''E. coli'' odor-free chassis=== | ||
| + | *Culture of MG1655 and MG1655Z1 strains- [https://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 30st July] | ||
*P1 phage stock preparation for the transduction of the ''Delta-tnaA::Kan'' - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/2 2nd July] | *P1 phage stock preparation for the transduction of the ''Delta-tnaA::Kan'' - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/2 2nd July] | ||
*P1 phage transduction using the stock prepared on July 2nd to MG1655 and MG1655Z1 strains - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/3 3rd July] | *P1 phage transduction using the stock prepared on July 2nd to MG1655 and MG1655Z1 strains - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/3 3rd July] | ||
| - | *Cultures of MG1655 and MG1655Z1 transductants - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/4 | + | *Cultures of MG1655 and MG1655Z1 transductants - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/4 4st July] |
*Transformations test of competent cells MG1655 and MG1655Z1 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/10 10th July] | *Transformations test of competent cells MG1655 and MG1655Z1 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/10 10th July] | ||
*Preparation of competent cells E coli, deletion of the antibiotic cassette in the odorless bacteria - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/11 11th July] | *Preparation of competent cells E coli, deletion of the antibiotic cassette in the odorless bacteria - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/11 11th July] | ||
| Line 28: | Line 24: | ||
*Preparation and Transformation of competent MG1655 and MG1655Z1 with BT340 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/28 28th July] | *Preparation and Transformation of competent MG1655 and MG1655Z1 with BT340 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/28 28th July] | ||
*PCR of MG1655 and MG1655Z1 with FTR-Apra-F and FTR-Apra-R - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/29 29th July] | *PCR of MG1655 and MG1655Z1 with FTR-Apra-F and FTR-Apra-R - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/29 29th July] | ||
| - | *Preparation and transformation of electrocompetent MG1655 and MG1655Z1 with plasmid BT340 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/30 | + | *Preparation and transformation of electrocompetent MG1655 and MG1655Z1 with plasmid BT340 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/30 30st July] |
*Clone isolation by streak of ''Delta-tnaA'' MG1655 and MG1655Z1 on LB dishes - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/1 1st August] | *Clone isolation by streak of ''Delta-tnaA'' MG1655 and MG1655Z1 on LB dishes - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/1 1st August] | ||
*Clone isolation by streak of ''Delta-tnaA'' MG1655 and MG1655Z1 on LB and kan dishes - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4 4th August] | *Clone isolation by streak of ''Delta-tnaA'' MG1655 and MG1655Z1 on LB and kan dishes - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4 4th August] | ||
*PCR verification of the final strains ''Delta-tnaA'' MG1655 and MG1655Z1 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/5 5th August] | *PCR verification of the final strains ''Delta-tnaA'' MG1655 and MG1655Z1 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/5 5th August] | ||
| - | *Final stock of MG1655 ''Delta-tnaA'' - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/6 | + | *Final stock of MG1655 ''Delta-tnaA'' - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/6 6st August] |
===The Lemon Scent=== | ===The Lemon Scent=== | ||
====Preparation==== | ====Preparation==== | ||
| - | * Rehydration of BioBricks BBa_J45014, BBa_K517003 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 | + | * Rehydration of BioBricks BBa_J45014, BBa_K517003 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 30th June] |
| - | * Preparation of electrocompetent DY330 and transformation via pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18#Preparation_of_electrocompetent_DY330_and_transformation_via_pJBEI6409 | + | * Preparation of electrocompetent DY330 and transformation via pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/18#Preparation_of_electrocompetent_DY330_and_transformation_via_pJBEI6409 18th July] |
| - | * Extraction of p cola plasmid DNA - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/22#Extraction_of_p_cola_plasmid_DNA | + | * Extraction of p cola plasmid DNA - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/22#Extraction_of_p_cola_plasmid_DNA 22nd July] |
| - | * Extraction and electrophoresis of BBa_K762100 with pSB1C3 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/24#Plasmid_DNA_Extraction_&_Electrophoresis | + | * Extraction and electrophoresis of BBa_K762100 with pSB1C3 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/24#Plasmid_DNA_Extraction_&_Electrophoresis 24th July] |
| - | * Gel electrophoresis of p cola - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/24#Gel_electrophoresis_of_p_cola | + | * Gel electrophoresis of p cola - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/24#Gel_electrophoresis_of_p_cola 24th July] |
====pPS1==== | ====pPS1==== | ||
| - | * PCR targeting with DY330 and pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/22#PCR_Targeting | + | * PCR targeting with DY330 and pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/22#PCR_Targeting 22nd July] |
| - | * Transformation of DY330 with pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/25#Transformation_of_electrocompetent_cells | + | * Transformation of DY330 with pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/25#Transformation_of_electrocompetent_cells 25th July] |
| - | * Liquid culture of DY330 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/28#Liquid_Culture | + | * Liquid culture of DY330 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/28#Liquid_Culture 28th July] |
| - | * Transformation of DY330 with pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/29#Transformation_of_electrocompetent_cells 29th | + | * Transformation of DY330 with pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/29#Transformation_of_electrocompetent_cells 29th July] |
| - | * Culture of DY330 + pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/31#D_-_Lemon_scent | + | * Culture of DY330 + pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/31#D_-_Lemon_scent 31st July] |
| - | * Transformation of DY330 with pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/1#New_Transformation_of_electrocompetent_cells | + | * Transformation of DY330 with pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/1#New_Transformation_of_electrocompetent_cells 1st July] |
| - | * Electroporation of DY330+pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4#Electroporation | + | * Electroporation of DY330+pJBEI-6409 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4#Electroporation 4th July] |
====pPS2-pPS3-pPS4==== | ====pPS2-pPS3-pPS4==== | ||
| - | *PCR of the differents genes or Biobrick [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/5#PCR_LS_PS_GS_Cad | + | *PCR of the differents genes or Biobrick [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/5#PCR_LS_PS_GS_Cad 5th Septembre] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/8#Checking_PCR 8th September] |
| - | * Cloning in Topo vector and transformation of competent E.coli [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/8#Cloning | + | * Cloning in Topo vector and transformation of competent E.coli [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/8#Cloning 8th September] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#ligation_of_PS_in_TOPO_vector_and_transformation 11th September] |
| - | * Checking of the cloning [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/9 | + | * Checking of the cloning [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/9 9th September] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/12 12th September] |
| - | * Plasmids extraction [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/10#Plasmid_extraction | + | * Plasmids extraction [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/10#Plasmid_extraction 10th Septembre] |
| - | * Digestion to have the insert [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_digestion | + | * Digestion to have the insert [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_digestion 11th September] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Digestion_of_PS_topo_plasmids 16th September] |
| - | * Ligation of the insert in the differents plasmids [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_ligation | + | * Ligation of the insert in the differents plasmids [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#LS_and_GS_ligation 11th September] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#Ligation 16th September] |
| - | * Transformation [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/15#Transformation_with_the_ligation_.28futur_pPS2.2F4.29 | + | * Transformation [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/15#Transformation_with_the_ligation_.28futur_pPS2.2F4.29 15th September] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/17#transformation 17th September] |
| - | * PCR on colony [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#PCR_Colony | + | * PCR on colony [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16#PCR_Colony 16th September] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/18 18th September] |
| - | * Checking of the insert sens [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/19#PCR_to_check_the_sens_of_the_insert | + | * Checking of the insert sens [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/19#PCR_to_check_the_sens_of_the_insert 19th September] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/23 here] and [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/24 24th September] |
| - | * Final stock and extraction of pPS3 and pPS4 [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/25 | + | * Final stock and extraction of pPS3 and pPS4 [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/25 25 September] |
====pPS5==== | ====pPS5==== | ||
| - | *Preparation of the pPS4 plasmid with SalI digestion [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion | + | *Preparation of the pPS4 plasmid with SalI digestion [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion 24th September] |
| - | *Ligation [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion | + | *Ligation [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/24#SalI_digestion 24 September] |
====PSBIC3==== | ====PSBIC3==== | ||
| - | *Preparation of the plasmids [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#Preparation_of_pSCBIC3_plasmid | + | *Preparation of the plasmids [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/11#Preparation_of_pSCBIC3_plasmid 11 September] |
| - | === | + | ===Salicylate Inducible Suppressing System=== |
| - | *Rehydration of BioBricks BBa_J61051 and BBa_K228001- [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/23# | + | *Rehydration of BioBricks BBa_J61051 and BBa_K228001- [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/23#The_Suppressing_Salicylate_System 23rd July] |
| - | *Transformation of competent E.coli cells - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/24# | + | *Transformation of competent E.coli cells - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/24#Salicylate_Inducible_Suppressing_System 24th July] |
| - | *Bacterial culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/25# | + | *Bacterial culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/25#Salicylate_Inducible_Suppressing_System 25th July] |
| - | *Plasmid DNA Purification - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/28# | + | *Plasmid DNA Purification - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/28#Salicylate_Inducible_Suppressing_System 28th July] |
| - | *Digestion to check & Electrophoresis - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/28# | + | *Digestion to check & Electrophoresis - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/28#Salicylate_Inducible_Suppressing_System 28th July] |
| - | *Main Digestion of BBa_J61051 and BBa_K228001 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/29# | + | *Main Digestion of BBa_J61051 and BBa_K228001 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/29#Salicylate_Inducible_Suppressing_System 29th July] |
| - | *Segregate Process - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/29# | + | *Segregate Process - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/29#Salicylate_Inducible_Suppressing_System 29th July] |
| - | *DNA purification gel agarose - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/30# | + | *DNA purification gel agarose - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/30#Salicylate_Inducible_Suppressing_System 30th July] |
| - | *Ligation - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/30# | + | *Ligation - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/30#Salicylate_Inducible_Suppressing_System 30th July] |
| - | *Transformation of competent E.coli cells - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/31# | + | *Transformation of competent E.coli cells - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/31#Salicylate_Inducible_Suppressing_System 31st July] |
| - | *Liquid Culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/1# | + | *Liquid Culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/1#Salicylate_Inducible_Suppressing_System 1st August] |
| - | *Plasmid DNA Purification - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4# | + | *Plasmid DNA Purification - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4#Salicylate_Inducible_Suppressing_System 4th August] |
| - | *Digestion to check & Electrophoresis - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4# | + | *Digestion to check & Electrophoresis - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4#Salicylate_Inducible_Suppressing_System 4th August] |
| - | *Final Stock '''BBa_K1372000''' - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4# | + | *Final Stock '''BBa_K1372000''' - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/4#Salicylate_Inducible_Suppressing_System 4th August] |
| - | *Liquid Culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/5# | + | *Liquid Culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/5#Salicylate_Inducible_Suppressing_System 5th August] |
| - | *Plasmid DNA Purification - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/6# | + | *Plasmid DNA Purification - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/6#Salicylate_Inducible_Suppressing_System 6th August] |
| - | *Digestion of BBa_K1372000 and BBa_B0015- [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/6# | + | *Digestion of BBa_K1372000 and BBa_B0015- [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/6#Salicylate_Inducible_Suppressing_System 6th August] |
| - | *Segregate Process - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/6# | + | *Segregate Process - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/6#Salicylate_Inducible_Suppressing_System 6th August] |
| - | *DNA purification gel agarose - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/7# | + | *DNA purification gel agarose - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/7#Salicylate_Inducible_Suppressing_System 7th August] |
| - | *Ligation - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/7# | + | *Ligation - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/7#Salicylate_Inducible_Suppressing_System 7th August] |
| - | *Transformation of competent E.coli cells - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/8# | + | *Transformation of competent E.coli cells - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/8#Salicylate_Inducible_Suppressing_System 8th August] |
| - | *Liquid Culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/11# | + | *Liquid Culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/11#Salicylate_Inducible_Suppressing_System 11th August] |
| - | *Plasmid DNA Purification - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/12# | + | *Plasmid DNA Purification - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/12#Salicylate_Inducible_Suppressing_System 12th August] |
| - | *Digestion to check & Electrophoresis - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/12# | + | *Digestion to check & Electrophoresis - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/12#Salicylate_Inducible_Suppressing_System 12th August] |
| - | *Final Stock '''BBa_K1372001''' - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/12# | + | *Final Stock '''BBa_K1372001''' - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/12#Salicylate_Inducible_Suppressing_System 12th August] |
| - | === | + | ===Construction of the Fusion Color Chromoprotein=== |
| + | * Plasmid DNA extraction and electrophoresis - [https://2014.igem.org/Team:Paris_Saclay/Notebook/July/24 24th July] | ||
| + | * PCR of the Chromoprotein - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/18 18th August] | ||
| + | * PCR Cloning in pGEMT easy - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/20 20th August] | ||
| + | * Transformation of DH5α with chromoprotein (in pGEMT easy) - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/21 21st August] | ||
| + | * Plasmid extraction - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/25 25th August] | ||
| + | * Plasmid extraction - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/26 26th August] | ||
| + | * Electrophoresis of pGEMT - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/27 27th August] | ||
| + | * Transformation of DH5α by pSB1C3 - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/28 28th August] | ||
| + | * Colony replication - [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/1 1st September] | ||
| + | * Liquid culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/2 2nd September] | ||
| + | * Plasmid extraction and gel electrophoresis - [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/3 3rd September] | ||
| + | * PCR - [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/5 5th September] | ||
| + | * Check sequence of the w6 clone - [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/8 8th September] | ||
| + | * PCR Transformation for stock of the w6 clone - [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/15 15th September] | ||
| + | * PCR colony w6 clone, Liquid culture - [https://2014.igem.org/Team:Paris_Saclay/Notebook/September/16 16th September] | ||
| + | |||
| + | ===The Lemon Shaping=== | ||
*Concentration Agar Test - [https://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 30th June] | *Concentration Agar Test - [https://2014.igem.org/Team:Paris_Saclay/Notebook/June/30 30th June] | ||
*Concentration Agar Test - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/5 5th August] | *Concentration Agar Test - [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/5 5th August] | ||
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*Coloration of agar [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/13 13th August] | *Coloration of agar [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/13 13th August] | ||
*Preparation of M63 medium [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/14 14th August] | *Preparation of M63 medium [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/14 14th August] | ||
| - | * Incubation of E. Coli with plasmid FNR RBS AmylCP in M63 [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/18 | + | * Incubation of E. Coli with plasmid FNR RBS AmylCP in M63 [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/18 18th August] |
*Transformation of odor free E. coli with plasmids coding Fluo Protein [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/27 27th August] | *Transformation of odor free E. coli with plasmids coding Fluo Protein [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/27 27th August] | ||
| - | * Results of Fluorescent Protein [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/29 | + | * Results of Fluorescent Protein [https://2014.igem.org/Team:Paris_Saclay/Notebook/August/29 29th August] |
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{{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
- PCR of the differents genes or Biobrick 5th Septembre and 8th September
- Cloning in Topo vector and transformation of competent E.coli 8th September and 11th September
- Checking of the cloning 9th September and 12th September
- Plasmids extraction 10th Septembre
- Digestion to have the insert 11th September and 16th September
- Ligation of the insert in the differents plasmids 11th September and 16th September
- Transformation 15th September and 17th September
- PCR on colony 16th September and 18th September
- Checking of the insert sens 19th September and here and 24th September
- Final stock and extraction of pPS3 and pPS4 25 September
pPS5
- Preparation of the pPS4 plasmid with SalI digestion 24th September
- Ligation 24 September
PSBIC3
- Preparation of the plasmids 11 September
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
Construction of the Fusion Color Chromoprotein
- Plasmid DNA extraction and electrophoresis - 24th July
- PCR of the Chromoprotein - 18th August
- PCR Cloning in pGEMT easy - 20th August
- Transformation of DH5α with chromoprotein (in pGEMT easy) - 21st August
- Plasmid extraction - 25th August
- Plasmid extraction - 26th August
- Electrophoresis of pGEMT - 27th August
- Transformation of DH5α by pSB1C3 - 28th August
- Colony replication - 1st September
- Liquid culture - 2nd September
- Plasmid extraction and gel electrophoresis - 3rd September
- PCR - 5th September
- Check sequence of the w6 clone - 8th September
- PCR Transformation for stock of the w6 clone - 15th September
- PCR colony w6 clone, Liquid culture - 16th September
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
