Week from September 28th, to October 4th, 2009

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September, 28th

September, 29th

• A14 induction test using IPTG

The part doens't show any induction!! To test what happened, we have inoculated A14 L-1 from glycerol stocks. tomorrow it will be diluted 1:100 in 2 falcons (5ml), one of wich will be induced 10nM.

• Inoculus of T9002 from glycerol stocks in 2 falons, one grown anaerobically and the other areobically. They have been induced with 3OC6HSL 10nM and the next morning the measure of GFP was about the half in the anaerobiotically grown culture respect to the aerobically grown one.

• Prepartion of 2 falcons 50ml containing 35ml LB+Amp+2%glucose and infected from glycerol stocks with B9 and F2620. They have been incubated overnight at 37°C statically, without shaking.

September, 30th

We watched B9 and F2620 grown overnight in anaerobic way. In B9 lots of foam has developed that we couldn't see in F2620. Is it for it fermented and made CO2? In the evening they have been pelletted and F2620's pellet was bigger than B9's one.

LIGATIONS:

• B7: A9-B5
• B9: F2620-B5
• B12: J118-B5
• B13: J106-B5
• A19: F2620-4Cm5

Inoculus of pnHAa and RBS32 for pH test, tomorrow will they will be diluted in 4 different LB, with different pH inoculus of A15-1, A15-3 F2620 from glycerol stocks for tomorrow lysis test. A15-3 has also been plated on LB+amp+agar2% to pick single colony for tomorrow test.

A14 inoculated yesterday has been diluted 1:100 into 2 falcons and induced with 3OC6HSLnM. Tomorrow we will see if the part works.

• Inoculus of 15ul from glycerol stock of B9 and F2620 in 2 cultures each, one closed ermetically (anaerobic) and the other a little open (aerobic). The cultures have been induced immediately with 3OC6HSL 10nM.

They have been incubated overnight at 37°C statically, without shaking.

October, 1st

October, 2nd

October, 3rd

October, 4th

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