About ZR Serum DNA Kit™
Note: This product is discontinued. For viral DNA extraction, please refer to ZR Viral DNA Kit™ (D3015/D3016) for the direct replacement product. For cell-free DNA extraction, please refer to Quick-cfDNA™ Serum & Plasma Kit (D4076) the direct replacement product.The ZR Serum DNA Kit™ is based on a state of the art, single buffer procedure for rapid DNA isolation from large volume serum and plasma samples. The product recovers genomic, mitochondria, and viral DNAs having typical sizes from 25 kb to 50 kb without RNA contamination. The uniquely formulated Genomic Lysis Buffer efficiently lyses cells, virus, and/or cellular particles. DNA/ZymoBead™ complexes are separated by centrifugation, and then washed to remove contaminants. Eluted, purified DNA is ideal for PCR and other sensitive analytical procedures.
|DNA Recovery||Scalable to compensate for smaller/larger sample sizes.
Not recommended for cell-free DNA isolation (see the Quick-cfDNA™ Serum & Plasma Kit)
|Equipment||Microcentrifuge or centrifuge (depending on sample volume).|
|DNA Size Limits||>1 kb|
|Sample Sources||Tissue, buccal cells, cells from culture, biological liquids, and blood from humans, mice, rabbits, etc.|
|DNA Purity||Purified, genomic DNA is recovered in water which is especially good for subsequent experiments. Mitochondrial and viral DNA will also be recovered.|
Ensure that the ZymoBeads™ are resuspended by vortexing. In a conical 50 ml tube add 4 volumes of Genomic Lysis Buffer to each volume of sample (4:1) and then add 10 µl of ZymoBeads™.
Example #1: 800 µl of plasma, add 3.2 ml of Genomic Lysis Buffer and 10 µl of ZymoBeads™.
Example #2: 5.0 ml of serum, add 20 ml of Genomic Lysis Buffer and 10 µl of ZymoBeads™.
Mix by placing sample on a rotator/rocker for 2 hours at room temperature or overnight at 0-4ºC.
|Step 3||Centrifuge tube for 1-2 minutes. Discard the supernatant.|
Add 500 µl of DNA Wash Buffer to the ZymoBeads™. Resuspend the pellet and then transfer mixture to a 1.5 ml microcentrifuge tube.
|Step 5||Centrifuge for 1 minute in a microcentrifuge. Discard the supernatant.|
Add another 500 µl of DNA Wash Buffer to the ZymoBeads™. Resuspend the pellet and centrifuge for 1 minute. Discard the supernatant.
|Step 7||Recentrifuge briefly and remove any residual wash buffer. Air-dry the pellet for 15 minutes.|
Add 10-35 µl DNA Elution Buffer or water to the ZymoBeads™ and resuspend by repeated pipetting.
|Step 9||Centrifuge at 10,000 x g for 1 minute.|
|Step 10||Collect the supernatant which contains the purified DNA. The DNA can be used immediately or stored at -20ºC for later use.|
Researchers studying circulating unmethylated and methylated preproinsulin (INS) DNA in type 1 diabetes (T1D) sought to test the differences between subjects with new-onset T1D and controls. DNA was extracted from serum using the ZR Serum DNA Kit™ and bisulfite converted with the EZ DNA Methylation Kit™ or the EZ DNA Methylation-Lightning Kit™. Droplet digital PCR was used to measure unmethylated and methylated serum levels. The data demonstrated that elevations in both unmethylated and methylated INS DNA occurs in new-onset T1D and that levels of these DNA species change during T1D evolution. The use of methylation levels as potential biomarkers for diabetes may prove to be a novel tool in the near future.