Team:Lyon-INSA-ENS/Realisation/Protocols/Fermentas

From 2011.igem.org

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-250 ng DNA ( if the concentration is unknown, use 5µL )<br/>
-250 ng DNA ( if the concentration is unknown, use 5µL )<br/>
-2.5µL (1X) or 5µL (2X) 10X tango buffer ( this depends on the enzymes used )<br/>
-2.5µL (1X) or 5µL (2X) 10X tango buffer ( this depends on the enzymes used )<br/>
-
-water to get a volume of 25mL after addition of the enzymes
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-water to get a volume of 25µL after addition of the enzymes
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     <p/>

Revision as of 09:56, 13 September 2011





Fermentas digestion





This protocol aims at cutting plasmids on specific restriction sites using restriction enzymes.





Procedure



1. In an eppendorf tube, add the following solutions in any order :
-250 ng DNA ( if the concentration is unknown, use 5µL )
-2.5µL (1X) or 5µL (2X) 10X tango buffer ( this depends on the enzymes used )
-water to get a volume of 25µL after addition of the enzymes


2. Add 1µL of each one of the desired enzymes.


3. Incubate at 37°C for 1h30


4. Incubate at 70°C for 10 mn to inactivate the restriction enzymes.








ENS assystem Biomérieux INSA INSA