protocol_496.txt

OmniPrep™ For High Quality Genomic DNA Extraction From Mouse Tail Tissue
Add 0.5-1cm, approximately 50-100mg, mouse tail in to a 1.5ml microcentrifuge tube with 500µl Genomic Lysis Buffer.
Add 10µl Proteinase K solution and incubate at 60°C for 3-4 hours to overnight.
Invert the tube periodically if possible.
Allow the sample to cool to room temperature.
Add 200µl chloroform and mix by inverting the tube several times.
Centrifuge for 10 minutes at 14,000xg and carefully remove the upper phase to a clean microcentrifuge tube.
Add 50µl DNA Stripping Solution to the sample and invert several times to mix.
Incubate the sample for 5-10 minutes at 60°C.
Add 100µl Precipitation Solution and mix by inverting the tube several times.
Centrifuge the sample at 14,000xg for 5 minutes.
Transfer the supernatant to a clean tube and precipitate the genomic DNA with 500µl isopropanol.
Invert the tubes 10 times to precipitate the DNA.
OPTIONAL: For increased DNA recovery, add 2µl Mussel Glycogen as a DNA carrier.
Centrifuge at 14,000xg for 5 minutes to pellet genomic DNA.
Remove the supernatant.
Add 700µl 70% ethanol to the tube and invert several times to wash the DNA pellet.
Centrifuge for 1 minute at 14,000xg.
Decant or pipette off the ethanol wash.  
Invert the tube on a clean absorbent surface for several minutes to allow any excess ethanol to drain away.
Add 50µl TE Buffer to the pellet.
Incubate at room temperature for at least 15 minutes to rehydrate.
OPTIONAL: 1µl LongLife™ RNase for every 100µl TE Buffer can be added at this stage.
Store DNA at 4°C, for long term storage store at -20°C or -80°C.