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Isolation of DNA from Agarose Gels (Paper Slurry Method)

This procedure isolates DNA from agarose gels by filtration through a filter-paper column. The column is made in a 500 µL tube from a slurry of filter paper in TE buffer.


Materials

 

Procedure

  1. Excise the DNA band from the surrounding gel with a clean razor blade; be sure to remove as little gel as possible.
  2. Dice the excised gel fragment into small pieces with the same razor; transfer onto filter column.
  3. Centrifuge for 10 minutes at highest speed (approximately 20,000g) in a microcentrifuge.
  4. Transfer eluent to a fresh tube; recentrifuge agarose
  5. Combine eluent with that from previous centrifugation
  6. Assemble a second spin column and transfer remaining agarose to the new column. Spin again.
  7. Combine all eluent fractions and concentrate via cold ethanol precipitation


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