Wednesday 6th August

Lab work

A - The chassis coli Odor free

Final stock of MG1655

by Romain

2 final stocks of the MG1655 made with 1ml of strain and 250µl glycerol 87%, in -80°C.

C - Salicylate Inducible Suppressing System

Plasmid DNA Purification

by Eugene and Fabio

• BBa_K1372000 Cl.1
• BBa_K1372000 Cl.2

From Liquid Culture made the 5th August

Protocol

Digestion

by Eugene and Fabio

In order to place the BBa_B0015 just after the BBa_K1372000, we will start the [http://parts.igem.org/Help:Assembly/3A_Assembly Assembly procedure] again.

• BioBrick BBa_K1372000 (Salicylate promoter + NahR + RNA suppressor) as Part A
• BioBrick BBa_B0015 (Double terminator) as Part B

The enzymes used were:

• EcoRI and SpeI for BBa_K1372000
• EcoRI and XbaI for BBa_B0015

TODO: Illustration of the process

BioBrick Assembly - Digest Reaction Protocol

Segregate Process by Electrophoresis

by Eugene and Fabio

1. BBa_K1372000 digested by EcoRI and SpeI
2. BBa_B0015 digested by EcoRI and XbaI

Results:

In progress

BioBrick Assembly - Segregate Process Protocol

D - Lemon scent

PCR Cleanup

by Pierre & Sean

BioBricks used: BBa_K762100 and BBa_K517003 (PCR perfomed on the 4th August)

Protocol

Gel electrophoresis

by Melanie

Plasmid used: p cola

Results inconclusive. Wrong enzyme was used to prepare PCR.

PCR digestion

by Pierre & Sean

Protocol

For each BioBrick (Bba_K517003 and BBa_K762100)mix the following into a microcentrifuge tube.

Then bain-marie at 37°C for one hour. Finally conduct PCR cleanup for both BioBricks, with 15µl of NE buffer.

Gel electrophoresis

(with Melanie)

Samples used: BBa_K517003 (2µl), BBa_K762100 (2µl), p cola(12µl)

Bibliography

By Hoang Vu

Still working on my mission: find more informations about limonene, pinene and citral A and B.I got now a couple of very interessant reviews about limonene and pinene.

Members present:

• Instructors and advisors: Alice.
• Students: Eugene, Fabio, Hoang Vu, Juliette, Melanie, Pierre, Romain, Sean and Terry.

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