Wednesday, June 01, 2005

Description
Small scale chromosomal DNA isolation from hair follicles. DNA is suitable for PCR amplification techniques.

Procedure

1. Take hair samples by pulling from just above the eye (do not cut!! we need the follicles!).
2. Cut of about 5-7 hair follicles into an Eppendorf tube.
3. Add 500ul TE (10mM Tris pH 8; 1mM EDTA) + 0.5% Tween 20.
4. Wash the follicles by vortexing.
5. Spin 1 min 13000rpm.
6. Aspirate sup as complete as possible.
7. Add 80ul of lysing buffer (0.3g Tris + 0.93g KCl + 1.25ml Tween-20 in total volume of 250ml) to the hair follicles and add 4ul of Proteinase-K (10mg/ml).
8. Incubate overnight (>6 hours) at 56 oC while gently rocking.
9. Heat the sample 10 minutes 94 oC.
10. Use 5ul of the supernatant in 50-100ul PCR reaction.


Yield of DNA is unknown. The washing step is crucial (especially with dirty hair). Please be aware of static electricity. Hairs will get everywhere!


Recipes
see above

Supplies
Eppendorf tubes

Tips
Please be aware of static electricity. Hairs will get everywhere!