DNA Isolation from hairs

Today...

I am planing to walk to the laboratory. Whuih....The weather so cold outside. It make me so lazy to walk, I remembered who nice weather in Indonesia... I prepare all the tools and foods. Here we go...!!!!

Today I am trying to isloate DNA from the hairs. It have never been done before, by my self or anyone in my laboratory. So its nice to do it...OK so what i have to prepare...
I prepare all the solutions below :
320 ul mix solutions :
- 50 uM Tris PH 7,5
- 10 uM EDTA
- 150 uM NaCl
- 1 % SDS
- 10 ul Proteinase K

And this how to perform Proteinase K solutions. Because when we order from the manufacturer. It is on powder form.
Proteinase K 10 ug/ul.
- 0,5 gr prot K (powder)
- 2,5 ul 20 % SDS
- add H2O total 50 ul and then incubate in 37 C 30 minutes.

OK let's we go how to perform it.
1. cut hair roots 0,5 cm 10-20 from hairs. Put on the mix solutions, incubate it in 55-56 C over night. (what for ?) it will be nice temp for prot K working.
2. take the mix tube from incubator.
3. let them in room temperature for 30-60 minutes.
4. add 100 ul 6 M NaCl.
5. spin down 15 minutes on high speed (13-14000 rpm)
6. take supernatant and move to new tube. You don't need pellet at this step, so trow away your pelet.
7. add 850 ul 96 % Ethanol or you can use absolute Ethanol to supernatant, shake smoothly and then spin down 1 minute on max speed.
8. you will get the pellet on this step and keep this pellet as your DNA remains in the pellet. So trow away your supernatant.
9. please keep remember position the tube when you do ultracentrifugation as pellet sometimes did not appear clearly.
10. add the pellet with 70% ethanol 350 ul and again spin down for 1 minutes on max speed.
11. hooray you get the pellet (sometime not clear)..trow away your supernatant and let them dry in RT for 15 minutes....
12 add 100 ul water or buffer and your DNA is ready for PCR...

OK enjoy it...