Team:University College London/Protocols/RestrictionEnzymeDigest1

From 2012.igem.org

(Difference between revisions)

Latest revision as of 01:36, 27 September 2012

Enzyme Digest Protocol 1

Step 1 - Thawing cells: Thaw all materials on ice

Step 2 - Adding Ingredient: Add the following ingredients to autoclaved/sterile eppendorf tubes

Component	Amount (ul) (one enzyme used)	Amount (ul) (two enzymes used)
dH20	2.5	1.5
Buffer 1x	1	1
DNA template	5	5
BSA	0.5	0.5
Enzyme 1	1	2
Enzyme 2	N/A	1

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: 2.5 hours

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

@@ Line 2: / Line 2: @@
 <html><img href="https://static.igem.org/mediawiki/2012/d/de/Ucl2012-protocols-logo-enzdig.png" /></html>
-== Enzyme Digest ==
+== Enzyme Digest Protocol 1 ==</noinclude>
-</noinclude>
 '''Step 1 - Thawing cells:''' Thaw all materials on ice
 '''Step 2 - Adding Ingredient:''' Add the following ingredients to autoclaved/sterile eppendorf tubes
-Insert Table
+{| class="wikitable"
+|-
+! Component !! Amount (ul) (one enzyme used) !! Amount (ul) (two enzymes used)
+|-
+| dH20 || 2.5 || 1.5
+|-
+| Buffer 1x || 1 || 1
+|-
+| DNA template|| 5|| 5
+|-
+| BSA || 0.5|| 0.5
+|-
+| Enzyme 1 || 1 || 2
+|-
+| Enzyme 2 || N/A|| 1
+|}
 '''Step 3 - Addition of BioBrick:''' Flick contents gently and centrifuge.