About ZR Plant/Seed DNA MicroPrep™
This product will be discontinued within the next 3 months (November 30th, 2016) or while supplies last. Please refer to ZR Plant/Seed DNA MiniPrep (D6020) for the direct replacement product. For a Microprep format, the following additional components can be purchased: Zymo-Spin IC (C1004-50) and Zymo-Spin IV-µHRC (C1022-50). Please refer to (D6020) protocol for more information.
The ZR Plant/Seed DNA MicroPrep™ is designed for the simple, rapid isolation of inhibitor-free, PCR-quality DNA from a variety of plant sample sources including leaves, stems, buds, flowers, fruit, seeds, etc. The procedure is easy and can be completed in minutes: plant samples are rapidly and efficiently lysed by bead beating with our state of the art, ultra-high density BashingBeads™. Polysaccharides, lipids, and polyphenols/tannins are removed from the DNA using our Fast-Spin column technology. The eluted DNA is filtered to remove polyphenolics making it ideal for downstream molecular-based applications including PCR, arrays, etc.
|DNA Recovery||Up to 5 μg total DNA is eluted into ≥ 10 μl DNA Elution Buffer per sample.|
|Processing Time||15 min|
|Equipment||Microcentrifuge, vortex, cell disruptor/pulverizer (optional)|
|DNA Size Limits||Capable of recovering genomic DNA up to and above 40 kb. In most instances, mitochondrial DNA and viral DNA (if present) will also be recovered.|
|Sample Sources||Up to 150 mg that include leaves, stems, buds, flowers, fruit, seeds, etc.|
|DNA Purity||High quality, inhibitor-free DNA is eluted with DNA Elution Buffer that is suitable for PCR amplification. A260/A280 > 1.8|
|DNA Yield||Typically 20-80 ng DNA/mg plant material is recovered.|
Comparison of DNA yields from various plant and seed samples using the ZR Plant/Seed DNA MiniPrep™. Equivalent amounts of plant materials were processed with equal volumes of eluted DNA analyzed in a 0.8% (w/v) agarose/ethidium bromide gel. M is a 1 kb DNA size marker (Zymo Research). Arabidopsis thaliana (1), juniper (2), corn kernel (3, 4), sunflower seed (5, 6).
Add up to 150 mg of finely cut plant or seed sample to a ZR BashingBead™ Lysis Tube. Add 750 µl Lysis Solution to the tube.
Secure in a bead beater fitted with a 2 ml tube holder assembly (e.g., Disruptor Genie™) and process at maximum speed for 10 minutes.
Processing times may be as little as 40 seconds when using high-speed cell disrupters (e.g., the portable Xpedition™ Sample Processor, page 6, FastPrepâ-24, or similar). See manufacturer’s literature for operating information.
Centrifuge the ZR BashingBead™ Lysis Tube in a microcentrifuge at ≥10,000 x g for 1 minute.
Transfer up to 400 µl supernatant to a Zymo-Spin™ IV Spin Filter (orange top) in a Collection Tube and centrifuge at 7,000 rpm (~7,000 x g) for 1 minute.
Add 1,200 µl of Plant/Seed DNA Binding Buffer to the filtrate in the Collection Tube from Step 4 and mix.
Transfer 800 µl of the mixture from Step 5 to a Zymo-Spin™ IC Column in a Collection Tube and centrifuge at 10,000 x g for 1 minute.
Discard the flow through from the Collection Tube and repeat Step 6.
Add 200 µl DNA Pre-Wash Buffer to the Zymo-Spin™ IC Column in a new Collection Tube and centrifuge at 10,000 x g for 1 minute.
Add 500 µl Plant/Seed DNA Wash Buffer to the Zymo-Spin™ IC Column and centrifuge at 10,000 x g for 1 minute.
Transfer the Zymo-Spin™ IC Column to a clean 1.5 ml microcentrifuge tube and add 20 µl (10 µl minimum) DNA Elution Buffer directly to the column matrix. Centrifuge at 10,000 x g for 30 seconds to elute the DNA.
Transfer the eluted DNA from Step 10 to a prepared Zymo-Spin™ IV-µHRC Spin Filter (yellow top) (see above) in a clean 1.5 ml microcentrifuge tube and centrifuge at exactly 8,000 x g for 1 minute. The filtered DNA is now suitable for PCR and other downstream applications.