About Quick-DNA™ 96 Kit
The Quick-DNA™ 96 Kit features a simple, high throughput (96-well) 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, buccal cells, cells from culture, and many biological liquid samples.
For processing, simply add the specially formulated Genomic Lysis Buffer to the samples, vortex, and transfer the mixtures to the wells of the supplied Silicon-A™ Plate. There is no need for organic denaturants or Proteinase K digestion because of the unique chemistries featured in the kit. Instead, the product yields high-quality, purified DNA in just minutes (see below). PCR inhibitors are effectively removed during the purification process. DNA purified using the Quick-DNA™ 96 Kit is suitable for PCR, nucleotide blotting, DNA sequencing, restriction endonuclease digestion, bisulfite conversion/methylation analysis, and other downstream applications.
|DNA Recovery||Up to 5 µg/well total DNA is eluted into ≥30 µl DNA Elution Buffer or water. Human whole blood will typically yield 3-7 µg DNA per 100 µl blood sampled. Mammalian tissues already homogenized yield: 1-3 µg DNA per mg skeletal, heart, and brain tissues and 3-5 µg DNA per mg liver, kidney and lung tissues.|
|Equipment||microcentrifuge, vortex, centrifuge w/ microplate carriers|
|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||Whole blood, plasma, or serum from humans, mice, rats, etc. Also, cells from culture, buccal cells, as well as a variety of biological liquids are effectively processed using this kit. Tissue already digested with Proteinase K or mechanically homogenized can also be processed.|
|DNA Purity||High-quality DNA is eluted with DNA Elution Buffer or water. DNA is especially well suited for PCR and other downstream applications. A260/A280>1.8|
|Product Detergent Tolerance||≤5% Triton X-100, ≤5% Tween-20, ≤5% Sarkosyl, ≤0.1% SDS.|
|Workflow Overview||Unique lysis buffer system omits the need for Proteinase K digestion for biological fluids and cell culture samples.|
Genomic DNA isolated from mouse tail snips using the Quick-DNA™ 96 Kit A total of 30 mouse tail snips were homogenized with Zymo Research's Squisher-8™ then processed using the Quick-DNA™ 96 Kit. About one third of the number of eluted DNAs was then separated in a 0.8% w/v agarose gel (shown in lanes 1 to 30).
Add 200 µl of Genomic Lysis Buffer to 50 µl of blood, serum, or plasma (4:1). Mix completely by vortexing 4-6 seconds, then let stand 5-10 minutes at room temperature.
Note: Add 200 µl Genomic Lysis Buffer to all samples < 50 µl. For samples larger than 50 µl, add a proportional amount (4:1) of Genomic Lysis Buffer (e.g., Add 400 µl Genomic Lysis Buffer to 100 µl blood).
Transfer the mixtures to the wells of a Silicon-A™ Plate on a Collection Plate. Centrifuge at ≥ 2,500 x g (5,000 x g max.) for 5 minutes.
Add 200 µl DNA Pre-Wash Buffer to each well and centrifuge at ≥ 2,500 x g for 5 minutes. Discard the flow through.
Add 300 µl of g-DNA Wash Buffer to each well and centrifuge at ≥ 2,500 x g for 5 minutes.
Transfer the Silicon-A™ Plate onto an Elution Plate. Add ≥30 µl DNA Elution Buffer or water to each well. Incubate 2-5 minutes at room temperature, then centrifuge at ≥ 2,500 x g for 5 minutes to elute the DNA. The eluted DNA can be used immediately for molecular based applications or stored ≤-20ºC for future use.