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Team:University College London/Protocols/RestrictionEnzymeDigest2
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| - | == Enzyme Digest == | + | == Enzyme Digest Protocol 2 == |
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'''Step 1 - Thawing cells:''' Thaw all materials on ice | '''Step 1 - Thawing cells:''' Thaw all materials on ice | ||
Revision as of 09:01, 1 August 2012
Enzyme Digest Protocol 2
Step 1 - Thawing cells: Thaw all materials on ice
Step 2 - Adding Ingredient: Add the following ingredients to autoclaved/sterile eppendorf tubes
Insert Table
Step 3 - Addition of BioBrick: Flick contents gently and centrifuge.
Step 4 - Centrifuge:
RPM: 14000
Time: 1 minute
Temperature: 18oC
Step 5 - Digest Program: Place the samples on a thermocycler under the following conditions:
RPM: 550
Time: 1 hour
Temperature: 37oC
Step 6 - Denaturing Enzymes: If you are not running the samples on a gel immediately, denature the restriction enzymes by running the samples on a thermocycler under the following conditions:
RPM: 550
Time: 25 minutes
Temperature: 65oC
Temperature: 18oC
