Team:Evry/Interlab Study/08-21-2014
From 2014.igem.org
(Difference between revisions)
Line 7: | Line 7: | ||
<p align="justify"> | <p align="justify"> | ||
- | <br/>Transformations of registry parts ( | + | <br/>Transformations of registry parts (BBa_K823012 and J23101) were performed to obtain colonies with plasmid containing parts for future amplifications. <br/> The transformation was performed on DH5 alpha ''E. coli'', as followed: </p> |
<ol> | <ol> | ||
<li> Remove E. coli competent tubes from -80°C and keep it on ice | <li> Remove E. coli competent tubes from -80°C and keep it on ice | ||
Line 15: | Line 15: | ||
<li> Incubate 2 minutes on ice | <li> Incubate 2 minutes on ice | ||
<li> Add 2 ml of LB medium and incubate 60 minutes at 37°C with an agitation at 200 rpm | <li> Add 2 ml of LB medium and incubate 60 minutes at 37°C with an agitation at 200 rpm | ||
- | <li> Plate 200 µl of | + | <li> Plate 200 µl of BBa_K823012 on a chloramphenicol LB agar plate or ampiciline for BBa_J23101 |
<li> Incubate plate overnight at 37°C | <li> Incubate plate overnight at 37°C | ||
</ol> | </ol> |
Revision as of 15:55, 28 August 2014
Transformations of registry parts (BBa_K823012 and J23101) were performed to obtain colonies with plasmid containing parts for future amplifications.
The transformation was performed on DH5 alpha ''E. coli'', as followed:
- Remove E. coli competent tubes from -80°C and keep it on ice
- Add 3 µl of template (here solubilized plasmids from the registry distribution kit) and mix gently
- Incubate 10 minutes on ice
- Perform an heat shock 30 seconds at 42°C
- Incubate 2 minutes on ice
- Add 2 ml of LB medium and incubate 60 minutes at 37°C with an agitation at 200 rpm
- Plate 200 µl of BBa_K823012 on a chloramphenicol LB agar plate or ampiciline for BBa_J23101
- Incubate plate overnight at 37°C