About ZR-96 Genomic DNA™ MagPrep
The ZR-96 Genomic DNA™ MagPrep features a streamlined, high throughput (96-well) procedure for 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.
PCR inhibitors are effectively removed during the purification process. DNA purified using the ZR-96 Genomic DNA™ MagPrep is suitable for PCR, nucleotide blotting, DNA sequencing, restriction endonuclease digestion, bisulfite conversion/methylation analysis, and other downstream applications.
|DNA Recovery||Up to 10 µg/well total DNA is eluted into ≥100 µl DNA Elution Buffer or water. Typical yields from samples are: 1-3 µg DNA per mg skeletal, heart, and brain tissues and 3-5 µg DNA per mg liver, kidney, and lung tissues.|
|Equipment||Automated liquid handler with heating element, plate shaker, and 96-well magnetic stand (not provided). The procedure can also be performed manually.|
|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||Solid tissues (e.g., tailsnips, earpunches, adipose tissue, etc.), whole blood, plasma, serum, buffy coat, lymphocytes, cultured cells, buccal cells, semen, tissues and other biological sources are effectively processed using this kit.|
|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.|
Proteinase K Digestion
The following is for the purification of up to 10 µg DNA from tissue or whole blood/cell suspensions.
1. To each sample (≤ 4 mg tissue or ≤ 70 µl biological fluid) in a well of a Collection Plate, add...
2X Digestion Buffer 70 µl
Proteinase K 10 µl
Sample X µl
*H2O to 150 µl
* For tissue samples, add 70 µl of H2O.
2. Mix and incubate at 55oC: 30 min for biological liquids and 1-3 hours for most tissues (until clarified).
3. Proceed to DNA Purification.
1. Dispense 450 µl of Genomic Lysis Buffer, 30 µl of MagBinding Beads, and mix well.
MagBinding Beads settle quickly, ensure that beads are kept in suspension while dispensing.
2. Vortex samples at ~1,300 rpm for 5-10 minutes.
3. Transfer the Collection Plate to a magnetic stand until beads have pelleted, then aspirate and discard supernatant.
4. Dispense 500 μl of Genomic Lysis Buffer and mix well.
5. Transfer the plate to the magnetic stand until beads pellet, then aspirate and discard the cleared supernatant.
6. Dispense 400 μl of DNA Pre-Wash Buffer and mix well.
7. Transfer the plate to the magnetic stand until beads pellet, then aspirate and discard the cleared supernatant
8. Dispense 400 μl of g-DNA Wash Buffer and mix well.
9. Transfer the plate to the magnetic stand until beads pellet, then aspirate and discard the cleared supernatant
10. Repeat the wash (steps 8-9) three times with 500 μl of g-DNA Wash Buffer.
11. Transfer the Collection Plate onto a heating element (65°C) until MagBinding Beads dry.
12. Dispense 100 μl of DNA Elution Buffer to each well of the Collection Plate and re-suspend beads by pipetting or vortexing. Place plate onto heating element and incubate for 10 minutes and then transfer the plate back onto the magnetic stand for 2-3 minutes until the beads pellet.
13. Transfer the supernatant (containing the eluted DNA) to a clean Elution Plate.
The DNA can be used immediately for molecular based applications or stored