Week from Semptember 7th, to September 13th, 2009

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September, 7th

In the morning we have prepared colture of A2 to stock in glycerol and we have prepared LB medium.

In the afternoon we have incubated in adequate medium:

• B9 (10 ml LB+Amp)
• B10 (10 ml LB+Amp)
• T9002 (10 ml LB + Amp)

to perform a qualitative test of ethanol production,

• A9pg (5 ml LB + Amp)
• A16 (5 ml LB + Amp)
• A8 (5 ml LB + Amp)
• A2 (5 ml LB + Amp)
• RBS33 (5 ml LB + Amp)

to perform an aTc induction test,

• A15 (25 ml LB + Amp)
• pSB3K3 (25 ml LB + Kan)
• A14-1 L (5 ml LB + Amp)

to miniprep tomorrow.

These coltures have been incubated at 37°C, 220 rpm overnight. Tomorrow, we will miniprep A14-1 L and midiprep A15 and 3K3 (pSB3K3). We perform midiprep on pSB3K3 because it is a low copy plasmid and on A15 because it is a lysis cassette, so for leackage only "few" cells survive. Glycerol stocks of A2-3 have been stored at -80°C, named A2-3 and A2-3 bis.The previous stock has been thrown away.

Preparation of a qualitative test to assay the presence of ethanol

We have also induced coltures B9 and B10 incubated yesterday morning at 37°C 220 rpm in flasks and grown overnight. In the morning the coltures have been diluited to reach the desired OD (0.03), then re-incubated for 4 hours and induced with 3OC6-HSL 10nM. The induced coltures have been incubated at 18.00 o'clock and will stay overnight. Flasks have been ermetically closed, to allow fermentation. This qualitative test is performed to assay the presence oh ethanol.

Tomorrow we will use the "Ethanol assay kit" to test the presence of ethanol!!

September, 8th

September, 9th

September, 10th

September, 11st

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