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1. Grind the tissue using a microtube micro-‐pestle until a uniform fine pulp is produced. Leave pestle in tube after grinding. 2. Transfer the macerated plant tissue to a ZR Bashing Bead Lysis Tube and add 650 μL Lysis Solution to the tube. Use additional 100 μL Lysis Solution to rinse the 1.5 mL Eppendorf tube and microtube micro-‐pestle used in step 1 and collect the rinse into the ZR Bashing Bead Lysis Tube. Cap tube tightly to prevent leakage. 3. Secure the ZR Bashing Bead Lysis Tubes in the Eppendorf ThermoMixer and process at maximum speed (2000 rpm) for 10 min. 4. Centrifuge the ZR Bashing Bead Lysis Tube in a microfuge at 13,000 rpm for 5 min. 5. Snap off the base of the Zymo-‐Spin IV Spin Filter (orange top) prior to use and insert into a new Collection Tube. Transfer up to 400 μL supernatant from step 4 to the prepared Zymo-‐Spin IV Spin Filter Collection Tube and centrifuge at 7,000 rpm for 1 min in a microfuge. 6. Add 1,200 μL of Plant/Seed DNA Binding Buffer to the filtrate in the Collection Tube from Step 4 and mix thoroughly by pipetting up and down with a 1 mL pipette tip. 7. Transfer 800 μL of the mixture from Step 6 to a Zymo-‐Spin IIC Column in a Collection Tube and centrifuge at 10,000 rpm in a microfuge. (The Zymo-‐Spin IIC Column has a maximum capacity of 800 μL). 8. Discard the flow through from the Collection Tube and repeat Step 7. Add 200 μL DNA Pre-‐Wash Buffer to the Zymo-‐Spin IIC Column in a new Collection Tube and centrifuge at 10,000 rpm for 1 min in an Eppendorf microfuge. 9. Add 500 μL Plant/Seed DNA Wash Buffer to the Zymo-‐Spin IIC Column and centrifuge at 10,000 rpm for 1 min in a microfuge. 10. Discard the flow-‐through in the Collection Tube and centrifuge again the Zymo-‐Spin IIC Column at 10,000 rpm for 1 min in a microfuge to completely remove the residual buffer. 11. Transfer the Zymo-‐Spin IIC Column to a clean 1.5 mL Eppendorf tube and discard the Collection Tube with flow-‐through. 12. Add 50 μL nuclease-‐free water directly to the middle of the column matrix of the Zymo-‐ Spin IIC Column and centrifuge at 10,000 rpm for 30 sec in a microfuge to elute the genomic DNA. 13. Remove the column and cap the tube. The DNA is now suitable for PCR and other downstream applications. 14. Store the DNA at -‐20oC until required. 35
