Protocol Still Needs Refining

Human DNA Extraction from Hair Follicle

DNA extraction for Southern Blot

1) Pluck a hair so that some root is removed from your head. The root is white/translucent in appearance.
Check to make sure you got some root and not all shaft.

2) Cut off most of the hair but keep the root (~ 5 mm). Be careful, sometimes the root will jump away when you cut the hair.

3) Incubate the root in 100 µl digestion buffer plus 6 µg of proteinase K for 1 hour at 55° C then 10 minutes at 95° C.

4) Use thermocycler program HAIR 1 - lid disabled for these temperatures and incubations times.

5) Do a phenol/chloroform extraction; then a chloroform extraction.

6) Add 1/10 volume sodium acetate and ethanol precipitate.

7) Spin for 10 minutes and then wash with 70% EtOH.

8) Resuspend DNA in 20 µl TE plus 2 µl RNase A.

Hair DNA Extraction Buffer - store at +4° C

• 1.86 g KCl
• 0.61 g Tris
• 0.25 g MgCl₂^.6H₂O
• 0.05g gelatin
• 2.25 ml NP40
• 2.25 ml Tween 20
• pH to 8.3 at RT°
• water to 500 ml and autoclave (will look white until it cools)

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