Monday 30th June

Lab work

A - The odor-free E. coli chassis

Culture of the supercompetents

by Mathias

• MG1655
• MG1655Z1

1ml of an overnight culture in 100ml of LB.

Rehydration of selected biobricks

by Juliette

• BBa_I9026 (rhII translational unit)
• BBa_K592025 (BB0034-amilCP)
• BBa_K786002 (sensory rhodopsin II & Tar sensitive to blue-green light)
• BBa_K786003 (sensory rhodopsin I & Tar sensitive to orange light)
• BBa_K778002
• BBa_K592011 (cjBlue green chromoprotein)(2 times)
• BBa_K592009 (amilCP blue chromoprotein)
• BBa_K103391 (Yellow chromoprotein and Lime yellow-green chromoprotein)(2 times)
• BBa_J45014 (lemon scent, alcohol acetyltransferase I)
• BBa_K517003 (lemon scent, β-pinene)

Transformations

by Maher & Mathias

We use the E. coli DH5α supercompetent cells provided by our advisors for the aforementioned Biobricks. Spread on petri dishes with the concentration 10^-1 and 10^-2.

Preparations

by Marie and Mathieu

• Aliquots of antibiotics

Put 0.203g of Chloramphenicol in 10ml of ethanol. Aliquots of 1ml. (Do the same for Kanamycin and Ampicillin)

by Romain

• Petri dishes

Add 250µl of antibiotics in 250ml of LB. Pour in Petri dishes.

by Arnaud

• Range of gelose with different concentration of agar

Add in 6 bottles of 500ml 2g of LB (lysogeny broth), and in each bottle add successively 1500mg (15mg/ml), 2000mg (20mg/ml), 2500mg (25mg/ml), 3000mg (30mg/ml), 3500mg (35mg/ml) and 4000mg (40mg/ml) of agar. And just before autoclaving complete each bottle with 100ml of reverse osmosis-purified water.

Members present:

• Instructors : Solenne.
• Students : Arnaud, Juliette, Maher, Marie, Mathias, Mathieu, Pierre, Romain.

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