Week from October 5th, to October 11st, 2009

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October, 5th

• Team Meeting

pH sensor

• Infection of 4 ml LB + Amp with 8 ul from glycerol stock of:
  • RBS33
  • A2
  • pNhaA
• Overnight incubation of these three cultures at 37°C, 220 rpm.

October, 6th

pH sensor

4th experiment

• We put 50 ul into 5 ml LB NaCl 250 mM + Amp pH 6,6 falcon for each of the overnight cultures RBS33 - A2 - pNhaA and incubated them for four five hours.
• OD measure of the three falcon with TECAN in order to make dilutions with a final volume of 3 ml and an OD equal to 0,02.
• We diluted:
  • RBS into:
    • LB NaCl 250 mM + Amp pH 5,5
    • LB NaCl 250 mM + Amp pH 6,6
    • LB NaCl 250 mM + Amp pH 7,5
    • LB NaCl 250 mM + Amp pH 8,5
  • pNhaA into:
    • LB NaCl 250 mM + Amp pH 5,5
    • LB NaCl 250 mM + Amp pH 6,6
    • LB NaCl 250 mM + Amp pH 7,5
    • LB NaCl 250 mM + Amp pH 8,5
  • A2 into:
    • LB NaCl 250 mM + Amp pH 5,5
    • LB NaCl 250 mM + Amp pH 6,6
    • LB NaCl 250 mM + Amp pH 7,5
    • LB NaCl 250 mM + Amp pH 8,5
• We started this 24 hours experiment that performed absorbance and fluorescence measures (on three different wells) for each sample every 5 minutes.

October, 7th

pH sensor

Qui risultati?

October, 8th

pH sensor

• Infection of 4 ml LB + Amp with 8 ul from glycerol stock of:
  • RBS33
  • A2
  • pNhaA
• Overnight incubation of the three cultures.

October, 9th

pH sensor

5th experiment (with this experiment we try to give E.coli a pH and sodium shock to see if something happens).

• We put 50 ul into 5 ml LB NaCl 70 mM + Amp pH 6,6 falcon for each of the overnight cultures RBS33 - A2 - pNhaA and incubated them for four four hours and a half.
• OD measure of the three falcon with TECAN in order to make dilutions with a final volume of 3 ml and an OD equal to 0,02.
• We diluted:
  • RBS into:
    • LB NaCl 600 mM + Amp pH 10
    • LB NaCl 600 mM + Amp pH 11,2
  • pNhaA into:
    • LB NaCl 600 mM + Amp pH 10
    • LB NaCl 600 mM + Amp pH 11,2
  • A2 into:
    • LB NaCl 600 mM + Amp pH 10
    • LB NaCl 600 mM + Amp pH 11,2
• We started this 6 hours experiment that performed absorbance and fluorescence measures (on three different wells) for each sample every 5 minutes.

October, 10th

pH sensor

Qui risultati?

October, 11th

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