Team:Tuebingen/Notebook/Protocols/restriction

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Preparative restriction digests

Reagents

min. 2 µg Plasmid DNA
2 µL of each restriction enzyme
10 µL 10x NE buffer 2
1 µL 100x BSA
to 100 µL H2O

 

Procedure

  1. After mixing the reagents in a 1.5 ml Eppendorf-tube: Incubation at 37 °C overnight.
  2. Heat inactivate the restriction enzymes at 80 °C for 20 min.
  3. Treat the plasmid backbones with Antarctic Phosphatase (NEB-protocol).
  4. Separation via gel electrophoresis and gel extraction.

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