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PLANT DNA PURIFICATION KIT DIRECTIONS
26-023 - Tissue Digestion
1. Mince up to 100 mg fresh/frozen
plant tissue
(up to 30 mg dried tissue)
2. Transfer to a clean 1.5 mL
microcentrifuge tube
(not provided)
3. Add 500 µL CTB Buffer. Vortex
vigorously to mix. Make sure to
disperse all clumps.
NOTE: Ensure βME is added to CTB
Buffer (20 µL βME/1 mL CTB).
Optional: Add 5 µL RNase. Let sit
at room temperature for 5 minutes.
Proceed to step 4.
4. Incubate at 65°C for 30 minutes.
Invert the samples twice
during incubation.
5. Add 700 μL chloroform/isoamyl alcohol (24:1). Vortex vigorously to mix.
6. Centrifuge at ≥10,000 x g for 5 minutes,
7. Transfer 300 μL aqueous phase (top) to a new microcentrifuge tube,
making sure not to disturb the pellet or transfer any debris.
8. Add 150 μL BB Buffer and 300 μL 100% ethanol. Vortex to obtain a
homogeneous mixture.
NOTE: A precipitate may form upon addition of ethanol; it will not interfere
with DNA isolation.
Plant DNA Purification Kit
26-023B - Tissue Homogenization
1. Weigh up to 100 mg of fresh/
frozen plant tissue (up to 30 mg
dried tissue) and transfer to the
2 mL tube containing 2.8mm
ceramic beads.
2. Add 500 µL of CTB buffer and
10 µL of Antifoam.
NOTE: Ensure βME is added to CTB
Buffer (20 µL βME/1 mL CTB).
3. Dissociate sample on a bead mill
at 3.5 – 4 m/s for 20 seconds or
with continuous vortexing for
10 minutes.
Optional: After homogenization,
add 5 µL RNase A. Let sit at room
temperature for 5 minutes.
Proceed to step 4.
4. Incubate at 65°C for 15 minutes.
Invert the samples twice
during incubation.
5
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