Template:SBB-Protocols Enz6

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Set up the following 10uL reaction in a PCR tube:
Set up the following 10uL reaction in a PCR tube:
<pre>
<pre>
-
4uL ddH2O
+
6.5 uL ddH2O
-
4uL Miniprepped plasmid
+
2uL Miniprepped plasmid
1uL 10x NEB Buffer 2
1uL 10x NEB Buffer 2
-
.5uL EcoRI
+
0.5uL EcoRI
-
.5uL BamHI (for parts >250bp) or XhoI (for parts <250bp)
+
0.5uL BamHI (for parts >250bp) or XhoI (for parts <250bp)
-
FOR Righty Methylated Parts use Eco/Xho!
+
</pre>
</pre>

Revision as of 16:58, 8 March 2012

Set up the following 10uL reaction in a PCR tube:

6.5 uL ddH2O
2uL Miniprepped plasmid
1uL 10x NEB Buffer 2
0.5uL EcoRI
0.5uL BamHI (for parts >250bp) or XhoI (for parts <250bp)
  • Incubate at 37 on the thermocycler for 30 minutes
  • Run an analytical gel
  • Take a picture of the gel
  • Calculate the expected fragment sizes
  • Are the calculated sizes consistent with the bands on the gel?
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