Duke University/9 June 2009

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June 9, 2009

Gel Extraction

  1. Weigh cut gel piece(s). Add at least 1 ul binding buffer per mg of gel.
  2. Place tube at 50-60°C for 10-15 min or until gel is completely dissolved. Vortex every few minutes.
  3. Add solution to spin column. Centrifuge at max speed for 30s. Discard filtrate.
  4. Add 300 ul binding buffer to column. Centrifuge for 30s. Discard filtrate.
  5. Add 500 ul wash buffer. Centrifuge for 30s. Discard filtrate.
  6. Repeat step 5.
  7. Centrifuge tubes for at least 2 min. Discard collection tube.
  8. Obtain 1.7 ml centrifuge tube. Add 40-50 ul elution buffer to spin column. Centrifuge for 1 min. Discard spin column.
  • E: Measured concentration of DNA fragments
  • R: Very low concentrations of fragments 1 and 3
  • C: Do PCR with fragments as template, greater volume to increase amount
  • E: PCR with fragments 1 and 3 as templates
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