About Quick-DNA™ Plant/Seed 96 Kit
Note: The ZR-96 Plant/Seed DNA Kit™ (D6021) has been updated to Quick-DNA™ Plant/Seed 96 Kit; all components and protocols are the same. Catalog numbers and names of components may have been modified but original buffer recipe remains the same.The Quick-DNA™ Plant/Seed 96 Kit 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 plate 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 ≥ 25 µl DNA Elution Buffer per sample.|
|Processing Time||50 min|
|Equipment||Centrifuge w/ microplate carriers, 96-well plate/block disruptor or pulverizer.|
|DNA Size Limits||Capable of recovering genomic DNA sized fragments from 100 bp to ≥ 40 kb. Typical fragment sizes range from 25 kb - 35 kb. If present, parasitic and viral DNA will also be recovered.|
|Sample Sources||Up to 80 mg of plant sample sources which include leaves, stems, buds, flowers, fruit, seeds, etc.|
|DNA Purity||High quality, inhibitor-free DNA is eluted with DNA Elution Buffer suitable for the amplification of bacterial, protist, and/or mammalian templates. Absorbance 260 nm/280 nm >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 Quick-DNA™ Plant/Seed 96 Kit. 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 80 mg of finely cut plant or seed sample to the tubes of a ZR BashingBead™ Lysis Rack. Add 400 µl Lysis Solution to each tube. Cap tubes tightly to prevent leakage.
Secure in a 96-well block/plate bead beater (e.g., 2010 GenoGrinder®) and process samples. Optimization of processing time/speed will be necessary for complete sample lysis.
Processing times may be as little as one minute when using high-speed bead beaters (e.g., 2000 GenoGrinder®, page 5). See manufacturer’s literature for specific operating information.
Centrifuge the ZR BashingBead™ Lysis Rack at ≥ 3,000 x g (5,000 x g max.) for 5 minutes.
Transfer up to 250 µl supernatant to each well of a 96-Well Block.
Add 750 µl of Plant/Seed DNA Binding Buffer to the supernatant in the 96-Well Block from Step 4. Cover completely with Cover Foil and mix thoroughly by vortexing for 2 minutes. Centrifuge the 96-Well Block at ≥ 3,000 x g (5,000 x g max.) for 5 minutes.
Remove or pierce foil and transfer 500 µl of each of the supernatants from Step 5 to the wells of a Silicon-A™ Plate on a Collection Plate. Centrifuge the assembly at ≥ 3,000 x g (5,000 x g max.) for 5 minutes.
Discard the flow through from the Collection Plate and repeat Step 6.
Add 200 µl DNA Pre-Wash Buffer to the wells of the Silicon-A™ Plate on the emptied Collection Plate and centrifuge the assembly at ≥ 3,000 x g for 5 minutes.
Add 500 µl Plant/Seed DNA Wash Buffer to the wells of the Silicon-A™ Plate on the Collection Plate and centrifuge the assembly at ≥ 3,000 x g for 5 minutes.
Transfer the Silicon-A™ Plate onto a clean Elution Plate and add 100 µl (50 µl minimum) DNA Elution Buffer directly to the matrices in the plate. Centrifuge the assembly at ≥ 3,000 x g for 5 minutes.
If fungi or bacterial cultures were sampled, the DNA is now suitable for PCR as well as other downstream applications.
Transfer the eluted DNA from Step 10 to a prepared Silicon-A™-HRC Plate mounted onto a clean Elution Plate. Centrifuge the assembly at exactly 3,500 x g for 3 minutes.
Eluted, ultra-pure DNA is now ready for use in your experiments, or the Elution Plate can be covered with Cover Foil for storage of the DNA.