Notebook

We constituted our team in November 2013 and we had meetings twice a month until April. Then we met every week. The Lab work started in the beginning of June.

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Planning

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 - Construction of the fusion protein (color)

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 - 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

E - Banana scent

F - 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