Abstract
Whole blood samples are critical for disease prediction and diagnosis. The emerging field of personalized medicine has significantly increased the demand of researcher’s usage of large volume of blood specimens from Biobanks. Our goal was to provide a scalable genomic DNA (gDNA) extraction method from whole blood suitable for downstream applications such as qPCR, microarray analysis and next generation sequencing (NGS).
Here we present an automated, high-quality, research-ready DNA extraction from large volume blood from biobanks. GenFind V3 DNA Isolation Kit integrated on KingFisher Duo system, provides scalable and consistent system for gDNA extraction from 2 mL of whole blood. GenFind V3 DNA Isolation Kit uses SPRI (Solid Phase Reverse Immobilization) paramagnetic bead technology to isolate genomic DNA from fresh or frozen whole blood and serum containing Citrate, EDTA or Heparin anticoagulants. The combined automation and chemistry platform processing 24 samples within 2 hours. A comparison between manual extraction and automated extraction was done with multiple donors. Quality metrics for gDNA yield, purity and integrity were accessed by NanoDrop and gDNA TapeStation. The gDNA yield from 2 mL whole blood was between 40-80 µg. Recovered gDNA exhibited high purity with A260/A280 ratios greater than 1.8 and A260/A230 ratios about 1.6. DNA integrity as measured by DNA integrity number (DIN) was above 9.0. The extract gDNA using GenFind V3 and KingFisher Duo system was suitable for downstream applications such as PCR, Microarray and NGS.