Dahlquist:DNA Extraction from Human Hair

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Overview

This protocol describes the steps necessary to obtain DNA from human hair for downstream PCR applications.

Recipes

5% Chelex

Makes 10 mL.

  • Weigh out 0.5 g of Chelex 100 (100-200 mesh, sodium form)
  • Add sterile MilliQ water, to make 10 mL of solution
  • Store at room temperature for one month.

Note that the Suenaga et al. article does not specify the buffer or pH of the solution. I found other sources online that say to resuspend the Chelex in sterile water.

Protocols

Prepare Hair

  • Pluck 2 hairs from head, making sure follicles are attached.
  • Tape down hair end (opposite of follicle end) and trim follicle end of hair to 1 cm length with clean scissors.
  • Place the trimmed hairs into 0.5 mL microcentrifuge tube.
  • Wash with 500 μL of 100% ethanol.
  • Air dry for 10 minutes in fume hood.

Chelex extraction

  • Add 200 μL of 5% Chelex 100 solution using cut tips and 2 μL of 50 μg/μL Proteinase K (100 μg) to microcentrifuge tube with hairs and mix well.
  • Incubate in 55°C water bath for at least 6-8 hours or overnight.
  • Vortex and incubate in a heat block set to 100°C for 8 minutes.
  • Spin in a microcentrifuge at 10,000 - 15,000 X g for 2-3 minutes.
  • Transfer 100 μL of the supernatant to another 0.5 mL microcentrifuge tube.
  • Use supernatant immediately for PCR or store at -20°C.

References

This protocol was adapted from: