DNA purification from
5-10 mg mouse tail tissue (Expected yield range 10-75 µg
- Chill a 1.5 ml tube containing 300 µl Cell Lysis Solution on
ice. Please note that the
solution will turn cloudy.
- Place 5 mm (5-10 mg) fresh or frozen mouse tail tissue (minced
if possible) into the
chilled Cell Lysis Solution.
- Add 1.5 µl proteinase K Solution (20 mg/ml) to the sample and
mix by inverting 25
times. Incubate at 55ºC overnight or until tissue has dissolved. If
tube periodically during the incubation.
RNase Treatment (optional)
- Add 1.5 µl RNase A Solution to the cell lysate.
- Mix sample by inverting the tube 25 times and incubate at 37ºC
for 15-60 minutes.
- Cool sample to room temperature.
- Add 100 µl Protein Precipitation Solution to the cell
- Vortex at high speed for 20 seconds to mix the Protein
uniformly with the cell lysate.
- Centrifuge at 13,000-16,000 x g for 3 minutes. The precipitated
proteins will form a
tight pellet. If the protein pellet is not visible, repeat step 3
followed by incubation on
ice for 5 minutes, then repeat Step 4.
- Pour the supernatant containing the DNA (leaving behind the
pellet) into a clean 1.5 ml microfuge tube containing 300 µl 100%
- Mix the sample by inverting gently 50 times.
- Centrifuge at 13,000-16,000 x g for 1 minute; the DNA will be
visible as a small white
- Pour off the supernatant and drain tube on clean absorbent
paper. Add 300 µl 70%
Ethanol and invert tube several times to wash the DNA pellet.
- Centrifuge at 13,000-16,000 x g for 1 minute. Carefully pour
off the ethanol. Pellet
may be loose so pour slowly and watch pellet.
- Invert and drain the tube on clean absorbent paper and allow to
air dry for 10-15
- Add 50 µl DNA Hydration Solution (50 µl will give a
concentration of 500 µg/ml if
the total yield is 25 µg DNA).
- Rehydrate DNA by incubating sample 1 hour at 65ºC and/or
overnight at room
temperature. If possible, tap tube periodically to aid in
dispersing the DNA.
- Store DNA at 4°C. For long-term storage, store at -20°C or
Copyright 2003 Gentra Systems, Inc.
Printed in USA •
2/03• 00708 Rev B