Difference between revisions of "Isolate genomic DNA from a single hair follicle"

Revision as of 13:51, 7 August 2012

Pluck a hair so that a large portion of root is removed from your head. For some people, the root will be white/translucent in appearance. People of African heritage will have roots that are dark. Regardless of the color, it will be sticky so you can test it by touching it to the bench top to see if it adheres. Check with someone else to make sure you got some root and not all shaft.
Put the hair into a PCR microfuge tube with the root at the bottom of the tube. 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.
Prepare hair DNA extraction buffer as follows: 500µL extraction buffer stock solution + 3 µL proteinase K stock (10mg/mL stock).
Use thermocycler program HAIR_1 - heated lid disabled. This program will incubate hair root in 100 µl extraction buffer + added proteinase K for 1 hour at 55˚C, then 10 minutes at 95˚C (what is the purpose of 95˚C?).

When the DNA extraction cools, vortex the tubes for 30 seconds and then set up a new 500 µl microfuge tube by adding the following:

PCR conditions:

END

Reference:
Campbell et al., 1997.

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 # Put the hair into a PCR microfuge tube with the root at the bottom of the tube. 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.
 # Prepare hair DNA extraction buffer as follows: 500µL extraction buffer stock solution + 3 µL proteinase K stock (10mg/mL stock).
-#  Incubate the root in 100 µl extraction buffer + proteinase K for 1 hour at 55˚C, then 10 minutes at 95˚C (what is the purpose of 95˚C?). Use thermocycler program HAIR_1 - '''heated lid disabled'''.
+# Use thermocycler program HAIR_1 - '''heated lid disabled'''. This program will incubate hair root in 100 µl extraction buffer + added proteinase K for 1 hour at 55˚C, then 10 minutes at 95˚C (what is the purpose of 95˚C?).