From 2014.igem.org
(Difference between revisions)
|
|
| (2 intermediate revisions not shown) |
| Line 1: |
Line 1: |
| - | <html>
| |
| - | <div id="cd-timeline" class="cd-container">
| |
| - | <div class="cd-timeline-block">
| |
| - | <div class="cd-timeline-img cd-picture">
| |
| - | <img src="img/cd-icon-picture.svg" alt="Picture">
| |
| - | </div> <!-- cd-timeline-img -->
| |
| | | | |
| - | <div class="cd-timeline-content">
| |
| - | <h2>Title</h2>
| |
| - | <p align="justify"> BBa_E0240 and BBa_I20260 are respectively into a PSB1C3 and a PSB1K3 vector. In order to select colonies, LB complemented with kanamycin or chloramphenicol are necessary. 200 ml of LB agar medium was prepared with 7 g of LB Agar powder(Sigma) into 200 ml of miliQ water. After complete dissolution, the solution was sterilized in the Tuttmauer 2540ML apparatus, STE 20 minutes at 121°C and EXT+DRY 15 minutes. 50 µl of kanamycin stock solution (25 mg/L) was added to 50 ml of LB agar to pour 2 plates. 150 µl of chloramphenicol stock solution was added in the 150 LB agar remaining ml to pour 6 plates.
| |
| - | <br/> The transformation was performed on DH5 alpha ''E. coli'', as followed: </p>
| |
| - | <ol>
| |
| - | <li> Remove E. coli competent tubes from -80°C and keep it on ice
| |
| - | <li> Add 1 µl of template (here solubilized plasmids from the registry distribution kit) and mix gently
| |
| - | <li> Incubate 10 minutes on ice
| |
| - | <li> Perform an heat shock 30 seconds at 42°C
| |
| - | <li> Incubate 2 minutes on ice
| |
| - | <li> Add 3 ml of LB medium and incubate 60 minutes at 37°C with an agitation at 200 rpm
| |
| - | <li> Plate 200 µl of BBa_E0240 on a chloramphenicol LB agar plate and BBa_I20260 on a Knamycin LB agar plate
| |
| - | <li> Incubate plate overnight at 37°C
| |
| - | </ol>
| |
| - | <span class="cd-date">Aug 15</span>
| |
| - | </div>
| |
| - | </div>
| |
| - | </div>
| |
| - | </html>
| |
Latest revision as of 12:29, 23 August 2014
"
