Week from September 21st, to September 27th, 2009

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September, 21st

September, 22nd

pH sensor

• LB NaCl 70 mM pH 5,5 - 6,6 - 7,5 - 8,5 + Amp preparation.
• 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.

September, 23rd

pH sensor

2nd experiment

• 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 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 70 mM + Amp pH 5,5
    • LB NaCl 70 mM + Amp pH 6,6
    • LB NaCl 70 mM + Amp pH 7,5
    • LB NaCl 70 mM + Amp pH 8,5
  • pNhaA into:
    • LB NaCl 70 mM + Amp pH 5,5
    • LB NaCl 70 mM + Amp pH 6,6
    • LB NaCl 70 mM + Amp pH 7,5
    • LB NaCl 70 mM + Amp pH 8,5
  • A2 into:
    • LB NaCl 70 mM + Amp pH 5,5
    • LB NaCl 70 mM + Amp pH 6,6
    • LB NaCl 70 mM + Amp pH 7,5
    • LB NaCl 70 mM + Amp pH 8,5
• We started this 6 hours experiment that performed absorbance and fluorescence measures (on three different wells) for each sample every 5 minutes.

September, 24th

pH sensor

Qui risultati?

September, 25th

pH sensor

• Preparation of LB NaCl 151 mM at pH 5,5 - 6,6 - 7,5 - 8,5.
• Preparation of LB NaCl 250 mM at pH 5,5 - 6,6 - 7,5 - 8,5.

September, 26th

September, 27th

pH sensor

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

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