OmniPrep™ For High Quality Genomic DNA Extraction From Blood
To <500µl whole blood, buffy coat, bone marrow or packed cells in a 2ml microfuge tube, add 0.75ml Nuclei Isolation Buffer.
Centrifuge at ~14,000g for 30 seconds to pellet the whole blood cells and nuclei.
Vortex to resuspend the pellet and add 0.75ml Nuclei Isolation Buffer.
Centrifuge at ~14,000g for 30 seconds to pellet the nuclei.
Incubate the sample at 55-60°C for 15 minutes.
Do not heat higher than 60°C.
OPTIONAL: For maximum DNA recovery, add 1µl Proteinase K solution for every 100µl Lysis Buffer and incubate at 60°C for 1-2 hours.
Invert the tube periodically each hour.
This step will digest hard to handle tissues and significantly improve the yield.
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.
Invert to mix and incubate at room temperature for 1 minute, invert at least twice during incubation.
Remove the supernatant containing lysed red blood cells, retaining the pellet.
Invert tube to mix and then incubate for 10 minutes at room temperature, inverting the tube every 1-2 minutes.
Remove the supernatant and retain the white nuclei pellet with 10-20µl supernatant.
Vortex to resuspend the pellet for improved nuclear lysis and add 500µl Genomic Lysis Buffer.
Mix by pipetting or vortexing at high speed for 10 seconds.
