Prepare DNA from Mouse Tail

Mouse tail lysis buffer

• 50 mM tris pH 8.0
• 100 mM EDTA
• 100 mM NaCl
• 1% SDS

Proteinase K

• stock at 20 mg/ml in H 2O
• aliquot and store at –20 oC

Use at 1 mg/ml = 1:20 dilution = 50 ul/ml

TE

• 10mM tris pH 8.0
• 1mM EDTA

Procedure

• Dilute proteinase K into mouse tail lysis buffer immediately before use.
• Add 400 ul above mixture to each tail.
• Incubate 55 o overnight or over weekend.

• Vortex well.
• Centrifuge 13,000 rpm for 5 minutes.
• Transfer supernatant to clean labeled microfuge tube.
• Add 800 ul isopropanol to sample.
• Invert a few times; then vortex. Should see DNA precipitate.
• Centrifuge 13K for 5 minutes.
• Dump off supernatant being careful not to lose pellet (should see visible white pellet on bottom of tube).
• Add 1 ml 70% EtOH (high quality).
• Vortex until pellet comes off bottom of tube.
• Centrifuge 13K for 5 minutes.
• Carefully dump off ethanol.
• Blot tube on paper towel and allow to drain.
• Dry completely.
• Add 400 ul TE to dissolve DNA. (best to let it dissolve overnight at 4 oC)
• Store at 4 oC, or at –20 oC for long term storage.