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Performance assessment of DNA sequencing platforms in the ABRF Next-Generation Sequencing Study



Justin Zook, Jonathan Foox, Christopher E. Mason


Assessing the reproducibility, accuracy and utility of massively parallel DNA sequencing platforms remains an ongoing challenge. Here the Association of Biomolecular Resource Facilities (ABRF) Next-Generation Sequencing Study benchmarks the performance of a set of sequencing instruments (HiSeq/NovaSeq/paired-end 2 × 250-bp chemistry, Ion S5/Proton, PacBio circular consensus sequencing (CCS), Oxford Nanopore Technologies PromethION/MinION, BGISEQ-500/MGISEQ-2000 and GS111) on human and bacterial reference DNA samples. Among short-read instruments, HiSeq 4000 and X10 provided the most consistent, highest genome coverage, while BGI/MGISEQ provided the lowest sequencing error rates. The long-read instrument PacBio CCS had the highest reference-based mapping rate and lowest non-mapping rate. The two long-read platforms PacBio CCS and PromethION/MinION showed the best sequence mapping in repeat-rich areas and across homopolymers. NovaSeq 6000 using 2 × 250-bp read chemistry was the most robust instrument for capturing known insertion/deletion events. This study serves as a benchmark for current genomics technologies, as well as a resource to inform experimental design and next-generation sequencing variant calling.
Nature Biotechnology


genomics, DNA sequencing, interlaboratory study, Next Generation Sequencing


Zook, J. , Foox, J. and Mason, C. (2021), Performance assessment of DNA sequencing platforms in the ABRF Next-Generation Sequencing Study, Nature Biotechnology, [online], (Accessed April 13, 2024)
Created September 9, 2021, Updated October 14, 2021