Jonesma Lab:Protocols:gelpurification

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Gel purification

  1. Run DNA out on a gel (Gel electrophoresis)
  2. Set a waterbath to 50°C
  3. Weigh a 1.5mL tube for each band to be cut
  4. Under a blue light (with an orange mat as a cutting board), cut out the bands needed (check sizes!) and put into individual tubes
  5. Reweigh tube with agarose cut out to find weight of agarose
  6. Add 3x volume of QG buffer (e.g. add 300μL buffer to a 100mg fragment)
  7. Incubate at 50°C for 10min, mixing occasionally
  8. Add to blue miniprep column (with clear plastic reservoir attached)- maximum volume 700μL
  9. Spin at 13 000rpm for 1min
  10. Empty reservoir and add remainder of dissolved agarose
  11. Spin at 13 000rpm for 1min
  12. Add 700μL PE buffer and incubate for 1min
  13. Spin at 13 000rpm for 1min
  14. Empty reservoir
  15. Spin at 13 000rpm for 1min
  16. Transfer blue column to new 1.5mL tube, labelled appropriately
  17. Add 40μL dH2O and incubate at room temperature for 1min
  18. Spin at 13 000rpm for 1min
  19. Keep tube with DNA, discard used blue column