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>
-
4.5 uL ddH2O
+
4uL ddH2O
4uL Miniprepped plasmid
4uL Miniprepped plasmid
1uL 10x NEB Buffer 2
1uL 10x NEB Buffer 2

Revision as of 17:22, 10 February 2010

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

4uL ddH2O
4uL 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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