Team:Cambridge/Protocols/Restriction Enzyme Digestion

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(Difference between revisions)
(Practice)
(Practice)
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* '''Master mix preparation'''
* '''Master mix preparation'''
: 2.0μl of DNA
: 2.0μl of DNA
-
: 0.5μl of each restriction enzyme (or a respective amount of water for the control with uncut plasmids)  
+
: 0.5μl of each restriction enzyme (or the respective amount of water for the control with uncut plasmids)  
-
: 0.1μl of BSA - acetylated Bovine Serum Albumin enhances the performance of restriction enzymes
+
: 0.1μl of BSA - ''acetylated Bovine Serum Albumin enhances the performance of restriction enzymes''
: 5.9μl of water
: 5.9μl of water
: 1.0μl of 2×NEBuffer
: 1.0μl of 2×NEBuffer
 +
 +
* '''Check for the compatibility of restriction enzymes chosen'''
 +
To check if the two selected restriction enzymes can perform effective catalysis in the same solution, go to the website of [http://www.neb.com New England Biolabs] → select
===Safety===
===Safety===

Revision as of 13:52, 15 July 2011

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Restriction Enzyme Digestion

protocol description

Theory

How it works

Practice

  • Master mix preparation
2.0μl of DNA
0.5μl of each restriction enzyme (or the respective amount of water for the control with uncut plasmids)
0.1μl of BSA - acetylated Bovine Serum Albumin enhances the performance of restriction enzymes
5.9μl of water
1.0μl of 2×NEBuffer
  • Check for the compatibility of restriction enzymes chosen

To check if the two selected restriction enzymes can perform effective catalysis in the same solution, go to the website of [http://www.neb.com New England Biolabs] → select

Safety

The safety implication of the procedure.