1. Cut 2-3mm tail tips and put in 1.5 ml tubes. 2. Add 600 microliters of 50mM NaOH. 3. Incubate at 100 C for 30-60 minutes until the tissue dissolves (for young tails <15 days it takes less than 30 min). I do it in a block heater. Be careful that the caps may pop, so I use those plastic lockers to keep them in place. 4. Cool down the tubes (I put them on ice for a minute). Add 50 microliters of 1M Tris.HCL (pH 8.0). Mix well by vortexing. 5. Spin in a microcentrifuge at top speed for 5 minutes. 6. (Optional) Transfer the supernatant to another tube and the DNA can be stored at 4 or -20 degrees. 7. Use one microliter (up to three microliter can be used) of the supernatant in a 50 microlitter reaction. I usually do 35 cycles and run 10 to 15 micrliters on a gel. |