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MOLECULAR BIOLOGY: WORKING WITH DNA
POLYMERASE CHAIN REACTION
MOUSE TAIL TIP DIGEST PROTOCOL: PREPARATION OF DNA
Mouse Tail Tip Digest Protocol: Preparation of DNA
1. Cut 20 to 30 mm of mouse tail from a mouse using a razor blade.
2. Place tails in labeled 1.5 ml microcentrifuge tubes.
3. Prepare digest mixture:
17 μl of Tail Tip Buffer
3 μl of 20 mg/ml Proteinase K
4. Add 20 μl of digest mixture from Step #3 into each tube containing tail tip.
5. Incubate tubes at 55°C for 1 to 2 hours.
6. After incubation, spin tubes for 10 seconds at top speed in a microcentrifuge.
7. Add 500 μl of ddH2O to each tube.
8. Boil samples for 5 minutes.
9. Spin samples briefly for 10 sec at top speed in a microcentrifuge and store tubes at 4°C for PCR analysis (see Protocol on PCR analysis of Tail Tip DNA).
Tail Tip Buffer 1 mM EDTA, pH 8.0
1% (w/v) SDS
50 mM Tris-HCl, pH 8.0
20 mM NaCl
Proteinase K 20 mg/ml Proteinase K in ddH2O
Store in aliquots at -20°C.
BioReagents and Chemicals
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1. Modified from the Hanahan Laboratory at UCSF.