Notebook

We constitute our team in November 2013 and we have meeting twice a month until April. Then we met every weeks. The Lab work started at the beginning of June.

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Planning

A - The chassis coli Odor free

• Culture of the supercompetent cells MG1655 and MG1655Z1 - 30th June
• Phages preparation for the transduction - 2nd July
• Phage transduction with the P1 phages in the strains MG1655 and MG1655Z1 - 3rd July
• Cultures of MG1655 and MG1655Z1 - 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 - 17th July
• Transformation of supercompetent cells MG1655 and MG1655Z1 - 18th July
• Results of the transformation - 21st July
• Preparation and transformation of competent MG1655 and MG1655Z1 - 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 BT340 - 30th July
• Stria of MG1655 and MG1655Z1 on LB dishes - 1st August
• Stria of MG1655 and MG1655Z1 on LB and kan dishes - 4th August
• PCR verification of the final strains MG1655 and MG1655Z1 - 5th August
• Final stock of MG1655 - 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 - 29th July
• Segregate Process - 29th July
• DNA extraction from 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_B0015 and BBa_K1372000 - 6th August
• Segregate Process - 6th August
• DNA extraction from 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 - 12th August

D - Lemon Scent

• 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

F - The lemon shaping

• Consentration Agar Test - 5th August