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Team:Evry/Interlab Study/08-21-2014
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| + | <h2>Contructions PSB1C3 with I20260, J23101-E1010 and K823012-E1010</h2> | ||
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| - | <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 | + | <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 <i>E. coli</i>, as followed: </p> |
<ol> | <ol> | ||
| - | <li> Remove E. coli competent tubes from -80°C and keep it on ice | + | <li> Remove <i>E. coli</i> competent tubes from -80°C and keep it on ice |
<li> Add 3 µl of template (here solubilized plasmids from the registry distribution kit) and mix gently | <li> Add 3 µl of template (here solubilized plasmids from the registry distribution kit) and mix gently | ||
<li> Incubate 10 minutes on ice | <li> Incubate 10 minutes on ice | ||
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<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 BBa_K823012 on a chloramphenicol LB agar plate | + | <li> Plate 200 µl of BBa_K823012 or BBa_J23101 on a chloramphenicol LB agar plate. |
<li> Incubate plate overnight at 37°C | <li> Incubate plate overnight at 37°C | ||
</ol> | </ol> | ||
Latest revision as of 12:13, 29 August 2014
Contructions PSB1C3 with I20260, J23101-E1010 and K823012-E1010
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 or BBa_J23101 on a chloramphenicol LB agar plate.
- Incubate plate overnight at 37°C
