using cheek cell dna

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    • #8026
      solong
      Participant

      Anyone with workable molecular lab protocols using dna from cheek cells.
      I work with it last semester but results were not so favorable.
      Any suggestion is appreciated.
      solong

    • #74923
      sdekivit
      Participant

      i did it a year ago. here’s the protocol

      1. vigorously scrape the inside of the cheek with a toothpick

      2. place the toothpick in a clean microcentrifuge tube and add 1 mL water. Vortex 10 seconds and spin in a centrifuge 45 seconds at 10.000 rpm.

      3. remove the toothpick and do not disturb the pellet! if pellet is distubed: centrifuge again.

      4. remove 770 uL water. do not disturb the pellet.

      5. add a little amount of chelex with a yellow tip.

      6. vortex for 10 sec.

      7. add 2 uL proteinase K and agitate.

      8. incubate at 56 C for 5 minutes.

      9. vortex for 10 seconds.

      10. incubate in boiling water bath for 8 minutes.

      11. vortex 10 seconds.

      12. spin in a microcentrifuge at maximum speed for 3 minutes.

      13. bottom = chelex and extraneous cellular matter, top layer = DNA. Sample = ready for PCR or may be stored at 4 C or frozen at -20 C. When reused after storage, thaw and repeat steps 12 and 13.

    • #74934
      solong
      Participant

      Thanx. I’ll try this

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