Team:UNIPV-Pavia/Notebook/Week4Sep
From 2009.igem.org
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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 + GFP (from now on it we'll be indicated only as pNhaA)
- Overnight incubation of the three culctures.
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
- RBS into:
- 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
September, 26th
September, 27th
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