Team:Grenoble/Biology/Protocols/Restriction
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Revision as of 09:33, 3 September 2012
Restriction enzyme digestion of DNA
Goal
Digest DNA in order to perform a ligation or to check a construction.Protocol
- Set up the water bath at 37°C.
- In an eppendorf tube, introduce :
- 2 µL of each restriction enzyme
- 10 µL of NEBuffer 2
- 1 µL of Bovine Serum Albumin (BSA).
- variable volume of template DNA
- to 50 µL of water
SAFETY AND USEFUL RECOMMANDATION Those products are not dangerous at first sight, but in case of spreading or in case of products on hands or on any parts on the body, it is recommended to wash this part and to dry it after. Be careful to use for each sample a different sterile cone. It is also important to well close tubes after putting the ingredient in. Moreover the DNA cannot resist until it is not incorporated in a plasmid and into a cell. Therefore there is no chance that there are any possible consequences because of a leakage in the environment.
- Put the eppendorf tube into the water bath.
- Incubate at 37°C for 15 minutes.