Mouse Tail DNA Extraction Protocol
1. Obtain tail biopsies from 2 to 4 week old mice: Hold mouse firmly at base of tail with one hand, with the other clean the tail with 70% alcohol and cut off 0.5 to 1.0 cm of the tail tip with a scalpel or single edge razor blade or sharp scissors. Clean the Put the tail tip into a 1.5 ml eppendorf tube.
2. Add 400 ul of TNES and 25 ul Proteinase K (10 mg/ml).
3. Incubate overnight (8-24 hr) at 55°C.
4. Add 166.7 ul 6M NaCl. Shake vigorously for 15 sec.
5. Microfuge (12,000-14,000 xg) for 5 min at room temperature.
6. Remove supernatant to new tube and add 1 volume cold 95% ethanol (EtOH).
7. Spool precipitated DNA with closed end capillary tube.
8. Rinse DNA pellet with 70% EtOH (dipping in a tube filled with 70% EtOH works well) and allow to air dry 5-10 min.
9. Resuspend in 100-500 ul of TE (10mM Tris, pH 8.0, 1mM EDTA). Volume depends empirically on pellet size.
10. Heat at 65°C for 10 minutes to aid dissolution of DNA.
11. Quantitate DNA and store at 4°C until needed.
TNES
10 mM Tris-HCl, pH 7.5
400 mM NaCl
100 mM EDTA
0.6% SDS
Or
100 mM Tris-HCl, pH 8.5,
5 mM EDTA,
0.2% SDS,
200 mM NaCl
"6M" NaCl Saturated salt solution stored at room temperature.
