Team:UNIPV-Pavia/Methods Materials/Digestion

From 2009.igem.org

(Difference between revisions)
(→BioBrick digestion with restriction enzymes)
Line 4: Line 4:
   <tr>
   <tr>
<td align="center">
<td align="center">
-
<img width="100%" height="180px" src="https://static.igem.org/mediawiki/2009/c/c2/UNIPV_title_MandM.jpg" ></img>
+
<img width="100%" height="180px" src="https://static.igem.org/mediawiki/2009/d/de/UNIPV_title_M%26M.jpg" ></img>
</td>
</td>
   </tr>
   </tr>

Revision as of 15:43, 17 October 2009

EthanolPVanimation.gif



Protocols


BioBrick digestion with restriction enzymes

(estimated time: 3 hours)

  • To open vectors:
    • a volume containing 1 µg of purified plasmid
    • 2 µl of buffer H
    • 1 µl of first enzyme
    • 1 µl of second enzyme
    • 20 µl final volume
    • incubate at 37°C for 3 hours
  • To excide fragments:
    • 20 µl of purified plasmid when <7 µg have been extracted. A volume containing 7 µg otherwise.
    • 2.5 µl of buffer H
    • 1 µl of first enzyme
    • 1 µl of second enzyme
    • 25 µl final volume
    • incubate at 37°C for 2 hours and 30 minutes






Roche restriction enzymes
Roche buffer H
Pre-warmed at 37°C bath
Cut and gel-extracted vector

ddH2O





Retrieved from "http://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/Digestion"