About ZymoBead™ Genomic DNA Kit
Note: This product will be discontinued within the next 3 months (November 30th, 2016) or while supplies last. Please refer to Quick-gDNA™ MiniPrep (D3024/3025) for the direct replacement product.
The ZymoBead™ Genomic DNA Kit is a simple procedure for the rapid isolation of total DNA (e.g., genomic, mitochondrial, viral) from a variety of biological sample sources. This product has been optimized for maximal recovery of ultra-pure DNA without RNA contamination and is compatible with whole blood (fresh or stored), serum, plasma, buffy coat, solid tissue, bone marrow and buccal cells, cells from culture, and many biological liquid samples. For processing, simply add the specially formulated Genomic Lysis Buffer to a sample in a 1.5 ml tube, add ZymoBeads™, vortex, then centrifuge. There is no need for organic denaturants or Proteinase K digestion because of the unique chemistries featured in the kit that yield high-quality, purified DNA in just minutes (see below). PCR inhibitors are effectively removed during the purification process. DNA purified using the ZymoBead™ Genomic DNA Kit is suitable for PCR, nucleotide blotting, DNA sequencing, endonuclease digestion, bisulfite conversion/methylation analysis, and other downstream applications.
|DNA Recovery||Typically, DNA is eluted into 35 μl DNA Elution Buffer or water for the standard procedure. Human whole blood will yield 3-7 μg DNA per 100 μl blood sampled. Mammalian tissues yield: 1-3 μg DNA per mg skeletal, heart, and brain tissues and 3-5 μg DNA|
|Processing Time||20 min|
|DNA Size Limits||Capable of recovering DNA up to and above 40 kb. In most instances, mitochondrial DNA and viral DNA (if present) will also be recovered.|
|Sample Sources||Whole blood, plasma, or serum from humans, mice, rats, etc. Also, tissue, cells from culture, buccal cells, as well as a variety of biological liquids are effectively processed using this kit.|
|DNA Purity||High-quality DNA is eluted with DNA Elution Buffer or water. DNA is well suited for PCR and other downstream applications. Typical absorption indices are A260/A280>1.8|
|ZymoBead™ Binding Capacity||~5 μg DNA per 10 μl ZymoBead™ slurry|
DNA isolation using the ZymoBead™ Genomic DNA Kit. Purifications were performed in duplicate for each sample and an equal volume of eluted DNA was loaded into each lane of a 0.8% (w/v) TAE/agarose/ethidium bromide gel. M is a 1 kb DNA ladder (Zymo Research)
Digested genomic DNA. Restriction endonuclease digestion of genomic DNA purified with the ZymoBead™ Genomic DNA Kit.
Ensure the ZymoBead™ slurry is fully resuspended by vortexing. In a 1.5 ml tube, add 200 µl of Genomic Lysis Buffer to 50 µl of blood, then add 10 µl ZymoBeads™. Mix by inversion, and then incubate at room temperature for 5 minutes. Centrifuge the tube at 1,500 x g for 1 minute. Carefully remove the supernatant without disturbing the bead pellet.
Add 200 µl of Genomic Lysis Buffer to the ZymoBeads™. Resuspend pellet by pipetting up and down. Centrifuge at 1,500 x g for 1 minute. Discard the supernatant.
Add 200 µl of DNA Pre-Wash Buffer to the ZymoBeads™. Resuspend the pellet, transfer to a new tube, and then centrifuge at 1,500 x g for 1 minute. Discard the supernatant.
Add 500 µl of g-DNA Wash Buffer to the ZymoBeads™. Resuspend the pellet and then centrifuge at 1,500 x g for 1 minute. Discard the supernatant. Recentrifuge briefly and remove any residual wash buffer.
Add ≥35 µl of Elution Buffer or water, resuspend pellet by pipetting up and down, and then centrifuge at 10,000 x g for 1 minute.
Collect the supernatant. The supernatant contains purified DNA that can be used immediately or stored (e.g.,-20ºC) for later use.