PAXgene Blood DNA Kit (25)

QIAGEN offers a PAXgene blood DNA kit that, together with PAXgene Blood DNA Tubes, provides an integrated, standardized system for the collection and stabilization of whole blood specimens and the isolation of high-quality genomic DNA. The collection tubes contain a proprietary blend of reagents that prevent blood coagulation and stabilize white blood cells; collected blood may be stored in the tubes for up to 14 days at room temperature (18–25°C), up to 28 days refrigerated (2–8°C), or frozen at −20°C or −70°C.

For purification, 8.5 ml blood samples are processed in single processing tubes prefilled with cell lysis buffer, with cell nuclei and mitochondria pelleted by centrifugation, washed, and resuspended in digestion buffer. Protein contaminants are removed by incubation with a protease, and DNA is precipitated in isopropanol, washed in 70% ethanol, and resuspended in resuspension buffer. This manual centrifugation-based protocol handles 8 samples in 60 minutes and yields 150–500 µg of genomic DNA in a 1 ml elution volume, using a salting-out method. The typical A260/A280 ratio is 1.7 to 1.9, with DNA molecules up to 200 kb in size and fragments of 50–150 kb predominating.

Purified DNA is free of detectable RNA contamination and contains both mitochondrial and genomic DNA, making it suitable for downstream applications including (RT-)PCR, (RT-)qPCR, NGS, genotyping, and microarray analysis. The system is intended for research use only and is designed for use with human blood samples. For storage, dissolved DNA should be kept at 2–8°C in Buffer BG4, or at –20°C for long-term storage, avoiding repeated freezing and thawing.