Dried blood spots for advanced DNA sequencing

Dried blood spots (DBS) are widely utilized in newborn screening, large population-based surveys, and biobanking due to their minimal blood volume requirements, ease of collection, and convenient transportation and storage. However, extracting high-quality DNA from DBS for next-generation sequencing (NGS) has posed significant challenges. This application note presents Revvity’s optimized workflow for targeted sequencing of DNA extracted from DBS, which addresses these challenges effectively.

The workflow begins with the DBS Puncher™ instrument, which punches samples into 96-well microtitration plates. DNA is then isolated using the chemagic™ DNA CS200 DNA Kit on the chemagic™ 360 instrument, capable of processing up to 1,000 samples per day. Central to this process is chemagen™ Technology, which employs an automated magnetic separation procedure to isolate high molecular weight (HMW) genomic DNA (gDNA) without denaturing or fragmenting nucleic acids. Quality control is ensured using the LabChip® GX Touch™ nucleic acid analyzer. The DNA is sheared to a target size of 150-200 bp, and libraries are prepared using Agilent® SureSelect® XT kits. The entire process is automated on the Sciclone® G3 NGSx workstation, and sequencing is performed on an Illumina® NovaSeq® sequencer.

The results demonstrate high-quality sequencing data with over 99% of reads mapped and low duplicate read percentages, confirming the effectiveness of Revvity’s workflow in producing pure, high-quality DNA suitable for NGS analysis. This optimized workflow facilitates the use of DBS in various research and biobanking applications, overcoming previous limitations in DNA extraction and sequencing.

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For research use only. Not for use in diagnostic procedures.