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Resources

TAIL DNA PROTOCOL    

 

Tail DNA isolation and purification

1. Add 500 ul Lysis buffer (100mM Tris HCl pH 8.5, 5mM EDTA, 0.2% SDS, 200 mM NaCl) to eppendorf tube containing tail biopsy. Add Proteinase K to a final concentration of 100ul/ml.

2. Incubate overnight on shaker at 62 degree C.

3. DNA lysis solution can be diluted 1/10 in water for most PCR applications.

4. To prepare DNA for Southerns and other sensitive applications: spin in centrifuge at maximum speed for 10 min.

5. Place 200-300 ul supernatant into new eppendorf tube

6. Add 500 ul isopropanol to supernatant

7. Shake for at least 5 minutes

8. Spin in centrifuge at maximum speed for 5 minutes

9. Pour off supernatant

10. Add 200 ul of 70% ethanol

11. Shake for at least 5 minutes

12. Spin in centrifuge at maximum speed for 5 minutes

13. Remove supernatant with pipetteman (do not pour off)

14. Dry pellet in warm chamber (37 degree C) for 15-30 minutes

15. Resuspend pellet in 100 ul TE pH 8.0 using wide bore tips

16. Put in 60 degree C for 5 hours to overnight

17. Store at 4 degree C