Team:UNIPV-Pavia/Methods Materials/Digestion

From 2009.igem.org

(Difference between revisions)
m
(→BioBrick digestion with restriction enzymes)
Line 49: Line 49:
**incubate at 37°C for 2 hours and 30 minutes
**incubate at 37°C for 2 hours and 30 minutes
<br><br><br><br>
<br><br><br><br>
-
<br></td><td align='center' valign='bottom' width='300px'>
+
<br></td><td class='digestion' align='center' valign='bottom' width='300px'>
-
<div align='center'>
+
<table class='in_digestion' border='1'>
-
<table border='1'>
+
<tr><td>
<tr><td>
*'''Roche restriction enzymes'''
*'''Roche restriction enzymes'''

Revision as of 17:23, 18 September 2009

EthanolPVanimation.gif



Protocols


BioBrick digestion with restriction enzymes

(estimated time: 3 hours)

</table>
Retrieved from "http://2009.igem.org/Team:UNIPV-Pavia/Methods_Materials/Digestion"
  • 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
</div>