From 2014.igem.org

Tuesday 11th August

Lab work

D - Lemon scent

PCR Clean-up of p cola and BBa_K517003

by Sean

PCR prepared on the 7th August.

Clean-up performed with the following protocol.

NB: in the present case we have 40 µl of each PCR result and we use 20 µl of elution buffer.

Protocol

Competents cells (CaCl2)

'by Melanie' protocol

stock of competents cells

Human Pratices

Art & Design

by Terry

Resumption of the preculture made yesterday, expressing blue chromoprotein. We will make 4 pieces of agar ( + control ) with different concentrations of bacteria in it.

Protocol :

- Preparation of a stock of 200ml agar gel at 20mg/ml. ( 4g of solid agar in 200ml of hot water )

- Sterilization of the mold with Ethanol 70%, all the work will be made in steril condition.

- Control of the Optic Density : 0.86

- 4 dilutions are prepared in total volume of 1ml :

• 1ml of preculture (100%)
• 500µl of LB and 500µl of preculture (50%)
• 900µl of LB and 100µl of preculture (10%)
• 990µl of LB and 10µl of preculture (1%)

- Introduction of 1ml of the different dilutions in the molds

- Introduction of 10ml of semi-liquid agar in each mold. Agar's temperature : 40°C

- The molds are securised with Aluminium.

- Direct incubation at 37°C

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