Team:Grenoble/Biology/Protocols/Restriction

From 2012.igem.org

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(Created page with "{{:Team:Grenoble/Templates/Biology}} <html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en"> <body id="Biology"> <div id="cadre"> <section> <h1>Restriction enzym...")
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                 <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr>
                 <tr><th><i><div class="petit"><center>SAFETY AND USEFUL RECOMMANDATION</center></div></i></th></tr>
                 <tr><td><div class="petit">Those products are not dangerous at first sight, but in case of spreading
                 <tr><td><div class="petit">Those products are not dangerous at first sight, but in case of spreading
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                 or in case of products on hands or on any parts on the body, it is recommended to wash this part and
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                 or contacts with any part of the body, it is recommended to wash this part and
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                 to dry it after.  Be careful to use for each sample a different sterile cone. It is also important to
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                 to dry it after.  Use a different pipette tip for each sample. It is also important to
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                 well close tubes after putting the ingredient in. Moreover the DNA cannot resist until it is not
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                 close tubes after putting the different reagents. DNA is more sensitive to its environment while it is not circulared in a plasmid and introduced
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                incorporated in a plasmid and into a cell. Therefore there is no chance that there are any possible
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                into a cell. Therefore, a spreading has very few chances to lead to any bad consequences.</div></td></tr>
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                consequences because of a leakage in the environment.</div></td></tr>
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         </table>
         </table>
     </center>
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Revision as of 14:45, 3 September 2012

iGEM Grenoble 2012

iGEM Grenoble iGEM
iGEM 2012
main page
Project

Restriction enzyme digestion of DNA

Goal

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

Protocol

  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

    SAFETY AND USEFUL RECOMMANDATION
    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.

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