"
|
SEPTEMBER: WEEK 1
September, 2ndE3-1C3-2 was purified in order to screen the part length.
R0040 in J61002-1C3, E3N-1C3 and J101-31N-1C3 were transformed in 200 μl of TOP10 competent cells.
September, 3rd
All the cultures were diluted 1:200 in M9 with the proper antibiotic. After two hours T9002-ENTERO and ENTERO-RBS were aliquoted in 96-well microplate and induced with known concentrations of 3OC6-HSL and with the supernatants collected on August, 31st.
2 μl of the purified plasmidic DNA were digested with 0.5 μl EcoRI and PstI restriction enzymes in a 25 μl final volume; a small size agarose gel was prepared.
The gel did not show the correct band.
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Team:UNIPV-Pavia/Calendar/September/week1
From 2011.igem.org
(Difference between revisions)
(Created page with "{{maincal}} <html> <h2 class="art-postheader"> SEPTEMBER: WEEK 1 </h2> <div class="cleared"></div> <div class="art-postcontent"> <p style="text-align:left;"> <p><a name="indic...") |
|||
| Line 32: | Line 32: | ||
<p> | <p> | ||
R0040 in J61002-1C3, E3N-1C3 and J101-31N-1C3 were transformed in 200 μl of TOP10 competent cells. | R0040 in J61002-1C3, E3N-1C3 and J101-31N-1C3 were transformed in 200 μl of TOP10 competent cells. | ||
| + | <br> | ||
| + | Inoculum of E24-2, E25-1, E26-2, E27-2 and RBS32 in M9 + Amp to collect supernatant after 3OC<small><sub>6</sub></small>-HSL supplementation. | ||
| + | <br> | ||
| + | Inoculum in M9 + Cm12.5 of T9002-ENTERO and ENTERO-RBS to measure 3OC<small><sub>6</sub></small>-HSL concentration in supernatants at different pHs collected on <a name="https://2011.igem.org/Team:UNIPV-Pavia/Calendar/August/week5#August.2C_31st">August, 31st</a>. | ||
| + | </p> | ||
| + | |||
| + | |||
| + | <div align="right"><small><a href="#indice" title="">^top</a></small></div> | ||
| + | |||
| + | <a name="September.2C_3rd"></a><h2> <span class="mw-headline">September, 3rd</span></h2> | ||
| + | <p> | ||
| + | All the cultures were diluted 1:200 in M9 with the proper antibiotic. After two hours T9002-ENTERO and ENTERO-RBS were aliquoted in 96-well microplate and induced with known concentrations of 3OC<small><sub>6</sub></small>-HSL and with the supernatants collected on <a href="https://2011.igem.org/Team:UNIPV-Pavia/Calendar/August/week5#August.2C_31st">August, 31st</a>. | ||
| + | <br> | ||
| + | E24-2, E25-1, E26-2, E27-2, RBS32 and sterile M9 were supplemented with 100 nM 3OC<small><sub>6</sub></small>-HSL. Supernatants were collected at t = 0 h, t = 7 h and t = 24 h from all of these tubes. | ||
| + | <br> | ||
| + | R0040 in J61002-1C3 and J101-31N-1C3 showed colonies while E3N-1C3 did not grow; two colonies were picked for each plate and inoculated in Lb + Cm34, 5 ml. | ||
| + | <br> | ||
| + | E3-1C3-2 was purified and quantified: | ||
| + | </p> | ||
| + | |||
| + | |||
| + | |||
| + | <p> | ||
| + | 2 μl of the purified plasmidic DNA were digested with 0.5 μl EcoRI and PstI restriction enzymes in a 25 μl final volume; a small size agarose gel was prepared. | ||
| + | </p> | ||
| + | |||
| + | |||
| + | <p> | ||
| + | The gel did not show the correct band. | ||
| + | <br> | ||
| + | 500 ml M9 and 1 l LB were prepared. | ||
</p> | </p> | ||
Revision as of 10:30, 4 September 2011
