"
Team:UNIPV-Pavia/Notebook/Week2Oct
From 2009.igem.org
(Difference between revisions)
m (→pH sensor) |
(→October, 8th) |
||
| Line 73: | Line 73: | ||
== <html><font class="dayw_style">October, 8th</font></html> == | == <html><font class="dayw_style">October, 8th</font></html> == | ||
| + | |||
| + | *Digestion screening for A20 (3 samples) and A21 (4 samples): all of them were positive! | ||
| + | |||
| + | *Over-day experiment on beta-gal activity using BioVision Lactose Assay kit: we diluted 1:100 the overnight cultures of B0033, A5 and V1022 in 30 ml of LB + Amp + 4.5% lactose for B0033 and A5 and in 30 ml of LB + 4.5% lactose without antibiotic for V1022. Then we incubated the three cultures (in 50 ml falcon tubes) at 37°C, 220 rpm; every 2 hours we measured the OD600 and the lactose using the commercial kit. | ||
| + | |||
| + | *All the streaked plates were grown and all the unshaken falcon tubes confirmed the phenotypes: B9-2, B9new-1, B9new-2 were cloudy, while B5new2-3 and F2620 were normal. This phenotype was not confirmed on the plates, which looked equal. | ||
| + | |||
===== pH sensor ===== | ===== pH sensor ===== | ||
*Infection of 4 ml LB + Amp with 8 ul from glycerol stock of: | *Infection of 4 ml LB + Amp with 8 ul from glycerol stock of: | ||
Revision as of 08:13, 15 October 2009
|
|
|
|
|
|
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
|
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
Week from October 5th, to October 11st, 2009
Previous Week
|
Next Week
|
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 at 37°C, 220 rpm.
- OD measure of the three falcons 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
- RBS into:
- 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
- Digestion screening for A20 (3 samples) and A21 (4 samples): all of them were positive!
- Over-day experiment on beta-gal activity using BioVision Lactose Assay kit: we diluted 1:100 the overnight cultures of B0033, A5 and V1022 in 30 ml of LB + Amp + 4.5% lactose for B0033 and A5 and in 30 ml of LB + 4.5% lactose without antibiotic for V1022. Then we incubated the three cultures (in 50 ml falcon tubes) at 37°C, 220 rpm; every 2 hours we measured the OD600 and the lactose using the commercial kit.
- All the streaked plates were grown and all the unshaken falcon tubes confirmed the phenotypes: B9-2, B9new-1, B9new-2 were cloudy, while B5new2-3 and F2620 were normal. This phenotype was not confirmed on the plates, which looked equal.
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, 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 at 37°C, 220 rpm.
- OD measure of the three falcons 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
- RBS into:
- 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
|
Previous Week
|
Next Week
|
