"
|
SEPTEMBER: WEEK 3
September, 12th
E31-1, E41N-1, E42-1, E43-3 and ENTERO4C5 were diluted 1:100 in 13 falcon tubes (4ml M9 + Cm12.5 pH = 6.0). After two hours they were induced with aTc (6 ng/ml, 8 ng/ml and 100 ng/ml). Supernatants were taken at the induction time, after 1 hour, 2 hours and 4 hours.
September, 13th
Supernatants collected on September, 12th were tested, measuring 3OC6-HSL concentration with T9002-ENTERO in two differents tests.
September, 14th
T9002-ENTERO, ENTERO-RBS were diluted 1:100 in M9 + Amp + Cm12.5 respectively in 18 ml and 1 ml. After two hours a test was performed on supernatants collected on September, 13th.
September, 15thSeptember, 16thSeptember, 17thSeptember, 18th
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Team:UNIPV-Pavia/Calendar/September/week3
From 2011.igem.org
(Difference between revisions)
| Line 22: | Line 22: | ||
<a name="September.2C_13th"></a><h2> <span class="mw-headline">September, 13th</span></h2> | <a name="September.2C_13th"></a><h2> <span class="mw-headline">September, 13th</span></h2> | ||
| + | <p> | ||
Supernatants collected on <a href="#September.2C_12th">September, 12th</a> were tested, measuring 3OC<sub><small>6</small></sub>-HSL concentration with T9002-ENTERO in two differents tests. | Supernatants collected on <a href="#September.2C_12th">September, 12th</a> were tested, measuring 3OC<sub><small>6</small></sub>-HSL concentration with T9002-ENTERO in two differents tests. | ||
<br> | <br> | ||
1:100 dilution of E37-2, E38-1, E39-1, E40-2 and ENTERO4C5 in 4ml M9 + Cm12.5 and two hours growth, after which cultures were induced with aTc; addition of 3OC<sub><small>6</small></sub>-HSL 100 nM after one hour and collection of supernatants at t = 0h, t = 1 h, t = 2 h and t = 4 h (time refers to 3OC<sub><small>6</small></sub>-HSL addition). | 1:100 dilution of E37-2, E38-1, E39-1, E40-2 and ENTERO4C5 in 4ml M9 + Cm12.5 and two hours growth, after which cultures were induced with aTc; addition of 3OC<sub><small>6</small></sub>-HSL 100 nM after one hour and collection of supernatants at t = 0h, t = 1 h, t = 2 h and t = 4 h (time refers to 3OC<sub><small>6</small></sub>-HSL addition). | ||
<br> | <br> | ||
| - | Inoculum of E44 for plasmid purification in 5 ml LB + Cm34; inoculum of T9002-ENTERO and ENTERO-RBS for 3OC<sub><small>6</small></sub>-HSL concentration measurement; ENTERO4C5 was inoculated in M9 + Cm12.5 at pH = 6.0 and pH = 7.0 in order to detect 3OC<sub><small>6</small></sub>-HSL degradation in | + | Inoculum of E44 for plasmid purification in 5 ml LB + Cm34; inoculum of T9002-ENTERO and ENTERO-RBS for 3OC<sub><small>6</small></sub>-HSL concentration measurement; ENTERO4C5 was inoculated in M9 + Cm12.5 at pH = 6.0 and pH = 7.0 in order to detect 3OC<sub><small>6</small></sub>-HSL degradation in cultures not expressing AiiA. |
</p> | </p> | ||
| Line 32: | Line 33: | ||
<a name="September.2C_14th"></a><h2> <span class="mw-headline">September, 14th</span></h2> | <a name="September.2C_14th"></a><h2> <span class="mw-headline">September, 14th</span></h2> | ||
| + | <p> | ||
| + | T9002-ENTERO, ENTERO-RBS were diluted 1:100 in M9 + Amp + Cm12.5 respectively in 18 ml and 1 ml. After two hours a test was performed on supernatants collected on <a href="#September.2C_13th">September, 13th</a>. | ||
| + | <br> | ||
| + | ENTERO4C5 was diluted in M9 + Cm12.5 at the proper pH; after growing for two hours at 37°C, 220 rpm, cultures were supplemented with 100 nM 3OC<sub><small>6</small></sub>-HSL and supernatants were taken at t = 0 h, t = 1 h, t = 2 h, t = 4 h and t = 8 h. | ||
| + | <br> | ||
| + | E44 was plasmid purified: | ||
| + | </p> | ||
| + | |||
Revision as of 14:27, 15 September 2011
