Team:TUDelft/15 July 2009

From 2009.igem.org

Lab Notebook

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15th July

Sriram

Cool. I got colonies in 11 plates out of 13 plates.

2 kanamycin plates with biobricks, mRFP1(E1010) and pBla (I14018), didn't contain any colonies and one other kanamycin plate with pLambclin promoter (I12006) has one colony.So the kanamycin containing biobricks must be re-transformed.

Checked for how to pick the colonies and grow in which tubes for biobrick amplification in order to assemble them. Found that the tubes were not autoclaved. So poured the medium in 24x5 ml tubes and given for autoclaving at 3PM.

Wow I got the autoclaved culture tubes at 4:30 PM, so with excitement I picked colonies (literally scooped) from 11 plates and grown in 11 antibiotic containing tubes (10 with 1xAmp and 1 with 1xKan) in shaker at 160 rpm at 37 °C.

Weenink

The BBa_K145280 had one colony. other ones didnt. It is not reccomendable to use this super fast protocol as it is very unreliable. 30 ml of LB 1x amp. was inoculated with this colony and put in a shaking incubator at 37 degrees and 160 rpm at 10:15.

I transformed DH5a competent cells with B0032, K145015, K145201 biobricks. I used the standard transformation protocol. Cells were incubated for 1h45m. at 37 degrees (accidentally not shaken) in 200µl SOC medium. Then 250µl of each transformation mix was plated on amp plates and put in the 37 degrees stove at 16:45.

Calin

Created a Matlab script based on the cascade delay ODEs, searched for proper parameters. Did some simulations with different parameter values. Began looking code needed for stability analysis.