(Difference between revisions)
Line 18: Line 18:
         <li>2 µL of each restriction enzyme</li>
         <li>2 µL of each restriction enzyme</li>
         <li>10 µL of NEBuffer 2</li>
         <li>10 µL of NEBuffer 2</li>
         <li>1 µL of Bovine Serum Albumin (BSA).</li>
         <li>1 µL of Bovine Serum Albumin (BSA)</li>
         <li>variable volume of template DNA</li>
         <li>variable volume of template DNA</li>
         <li>to 50 µL of water</li>
         <li>to 50 µL of water</li>

Revision as of 09:30, 5 September 2012

iGEM Grenoble 2012


Restriction enzyme digestion of DNA


Digest DNA in order to perform a ligation or to check a construction.


  1. Set up the water bath at 37°C.

  2. 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

    Those products are not dangerous at first sight, but in case of spreading or contacts with any part of the body, it is recommended to wash this part and to dry it after. Use a different pipette tip for each sample. It is also important to close tubes after putting the different reagents. DNA is more sensitive to its environment while it is not circulared in a plasmid and introduced into a cell. Therefore, a spreading has very few chances to lead to any bad consequences.

  3. Put the eppendorf tube into the water bath.

  4. Incubate at 37°C for 15 minutes.