Preparation of Genomic DNA from mouse tails
This protocol is for high quality DNA for Southern analysis for screening potential hetero- or homozygotes.
1. Cut 1 cm of mouse tail and place in 1.5 ml eppendorf tube
2. Add 500 ul of 50 mM Tris (pH 8), 100 mM EDTA, and 0.5% SDS
3. Add 25 ul of 10 mg/ml of proteinase K and mix well
4. Incubate at 55 C in a waterbath overnight
5. Next day add 500 ul of phenol and rotate for 20 min. at room temp.
6. Centrifuge at 14,000 rpm for 3 min at room temp
7. Take off supernatant and repeat steps 5-6
8. Take supernatant and add 500 ul of phenol/chloroform (1:1) and rotate for 20 min
9. Spin at 14,000 for 5 min
10. Transfer the top aqueous layer to a clean tube.
11. Add 50 ul of 3 M sodium acetate (pH 6) and 500 ul of 100 % ethanol. Mix well, (you should be able to see DNA precipitate)
12. Remove as much of ethanol supernatant as possible
13. Wash pellet with 1 ml of 70% ethanol 2X
14. Spin in microcentrifuge at 14,000 for 1 min
15. Aspirate supernatant carefully using a pipette tip, dry with speed vac if available
16. Add 100 ul of water and store at 4ƒ C overnight to dissolve.
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