OmniPrep™ For High Quality Genomic DNA Extraction From Plant Tissue ( Fresh or Frozen )
Most plant tissues are best prepared by freezing in liquid nitrogen .
Grind samples in liquid nitrogen to a fine powder and quickly add to an appropriate volume of Lysis Buffer .
Add 50 - 100mg finely ground dried tissue , frozen tissue or fresh leave tissue to a microcentrifuge tube containing 500µl Genomic Lysis Buffer .
If ground , vortex for 5 seconds ; if unground , homogenize the sample with a microfuge pestle until a homogenous suspension is acquired , approximately 30 - 60 strokes .
Incubate the sample at 65°C for 60 minutes with periodic inversions .
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 .
