Isolate genomic DNA from a single hair follicle
- 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. For others, the roots may be as black as the hair shaft. 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. This is the most important step - collecting enough soft tissue.
- Put 1-10 hairs into a PCR microfuge tube with the root(s) 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.
- The DNA extraction buffer was prepared as follows: 500µL extraction buffer stock solution + 3 µL proteinase K stock (10mg/mL stock). This has been done for you. Add 100 µL of hair extraction buffer to your hair follicle.
- Use thermocycler machine, 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 extracted gDNA mixture cools, vortex the tube for 30 seconds to make sure the DNA is well mixed. You can store this DNA at -20˚C if you are not going to use the DNA right away.
- Some users will amplify their TAS2R38 alleles TAS2R38_PCR_amplification.
Campbell et al., 1997.