Labwork

Planning

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

B - The Lemon Scent

• 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
• PCR targeting with DY330 and pJBEI-6409 - 22nd July
• Extraction and electrophoresis of BBa_K762100 with pSB1C3 - 24th July
• Gel electrophoresis of p cola - 24th July
• Transformation of DY330 with pJBEI-6409 - 25th July
• Liquid culture of DY330 - 28th July
• Transformation of DY330 with pJBEI-6409 - 29th July
• PCR of BBa_K762100 - 29th July
• PCR of BBa_K762100 - 30th July
• Culture of DY330 + pJBEI-6409 - 31st July
• Transformation of DY330 with pJBEI-6409 - 1st August
• Electroporation of DY330+pJBEI-6409 - 4th August
• PCR of BBa_517003 and BBa_K762100 - 4th August
• Gel electrophoresis of BBa_K517003 and BBa_K762100 - 5th August
• PCR cleanup of BBa_K517003 and BBa_K762100 - 6th August
• PCR of BBa_517003 and BBa_K762100 - 7th August
• PCR Clean-up of BBa_K762100 - 11th August
• PCR Clean-up of p cola and BBa_K517003 - 12th August
• Gel electrophoresis of p cola, BBa_K762100 and BBa_K517003 - 12th August
• Gel electrophoresis of BBa_K762100 - 13th August
• Cloning BBa_K517003 and p cola in pGEMTeasy - 13th August
• PCR of BBa_K517003 - 18th August
• PCR of BBa_K762100 - 18th August
• Transformation of DH5α with CAD and chromoprotein - 21st August
• Extraction of pPS3 and pPS4 - 26th August

C - 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

D - 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