Difference between revisions of "Team:Aix-Marseille/Protocols"
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<button type="button" class="btn btn-info" data-toggle="collapse" data-target="#demo1">Transformation</button> | <button type="button" class="btn btn-info" data-toggle="collapse" data-target="#demo1">Transformation</button> | ||
<div id="demo1" class="collapse"> | <div id="demo1" class="collapse"> | ||
| − | + | Plasmids transformation :<br /> | |
| − | + | ||
Add 20 ng of plasmid to 100 µL of competent cells thawed in ice<br /> | Add 20 ng of plasmid to 100 µL of competent cells thawed in ice<br /> | ||
Incubate 30-45 min in ice<br /> | Incubate 30-45 min in ice<br /> | ||
| Line 163: | Line 162: | ||
Suspend the pellet <br /> | Suspend the pellet <br /> | ||
Spread 150 µL on LB limp (with antibiotic)<br /> | Spread 150 µL on LB limp (with antibiotic)<br /> | ||
| − | To make negative control, follow the same procedure but without add plasmids and spread 300 µL | + | To make negative control, follow the same procedure but without add plasmids and spread 300 µL |
</div> | </div> | ||
</div> | </div> | ||
Revision as of 17:18, 15 September 2015
Protocols
Plasmids transformation :
Add 20 ng of plasmid to 100 µL of competent cells thawed in ice
Incubate 30-45 min in ice
Thermal shock : put tubes in the Thermomixer at 42°C during 2 min
Incubate 5 min in ice
Add 900 µL of LB
Incubate 1 hour at 37°C with agitation
Spread 100 µL on LB limp (with antibiotic)
To make negative control, follow the same procedure but without add plasmids and spread 300 µL
Ligation transformation:
Add 20 ng of ligation’s product to 100 µL of competent cells thawed in ice
Incubate 30-45 min in ice
Thermal shock : put tubes in the Thermomixer at 42°C during 2 min
Incubate 5 min in ice
Add 900 µL of LB
Incubate 1 hour at 37°C with agitation
Centrifuge 5 min at 5000 rpm
Eliminate 850 µL of medium
Suspend the pellet
Spread 150 µL on LB limp (with antibiotic)
To make negative control, follow the same procedure but without add plasmids and spread 300 µL
Add 20 ng of plasmid to 100 µL of competent cells thawed in ice
Incubate 30-45 min in ice
Thermal shock : put tubes in the Thermomixer at 42°C during 2 min
Incubate 5 min in ice
Add 900 µL of LB
Incubate 1 hour at 37°C with agitation
Spread 100 µL on LB limp (with antibiotic)
To make negative control, follow the same procedure but without add plasmids and spread 300 µL
Ligation transformation:
Add 20 ng of ligation’s product to 100 µL of competent cells thawed in ice
Incubate 30-45 min in ice
Thermal shock : put tubes in the Thermomixer at 42°C during 2 min
Incubate 5 min in ice
Add 900 µL of LB
Incubate 1 hour at 37°C with agitation
Centrifuge 5 min at 5000 rpm
Eliminate 850 µL of medium
Suspend the pellet
Spread 150 µL on LB limp (with antibiotic)
To make negative control, follow the same procedure but without add plasmids and spread 300 µL
Transformation
TITTLE
TEXTE
TEXTE
TEXTE
TEXTE
TEXTE
