Difference between revisions of "Isolate genomic DNA from a single hair follicle"
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Macampbell (talk | contribs) (Created page with '== DNA extraction == # 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 Afric…') |
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# 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: | # 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: | ||
Revision as of 21:05, 6 July 2012
DNA extraction
- 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.
- Incubate the root in 100 µl digestion buffer (which contains 6 µg of proteinase K) for 1 hour at 55˚C, then 10 minutes at 95˚C (what is the purpose of this step?). Use thermocycler program HAIR 1 - lid disabled.
PCR Mixture
- 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:
| Reagent | Volume |
|---|---|
| extracted DNA | 10 µL |
| 2X Green GoTaq Mix | 12 µL |
| Forward Primer | 1 µL |
| Reverse Primer | 1 µL |
| Final Volume | 24 µL |
Reference:
Campbell et al., 1997.
