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.

评估大规模并行 DNA 测序平台的再现性、准确性和实用性仍然是一个持续的挑战。在这里，生物分子资源设施协会 (ABRF) 下一代测序研究对一组测序仪器的性能进行了基准测试（HiSeq/NovaSeq/双端 2 × 250 bp 化学、Ion S5/Proton、PacBio 循环一致性测序 (CCS) ）、Oxford Nanopore Technologies PromethION/MinION、BGISEQ-500/MGISEQ-2000 和 GS111）对人类和细菌参考 DNA 样本进行分析。在短读长仪器中，HiSeq 4000和X10提供了最一致、最高的基因组覆盖率，而BGI/MGISEQ提供了最低的测序错误率。长读仪器 PacBio CCS 具有最高的基于参考的映射率和最低的非映射率。两个长读长平台 PacBio CCS 和 PromethION/MinION 在重复序列丰富的区域和同聚物中显示出最佳的序列图谱。 NovaSeq 6000 使用 2 × 250 bp 读取化学，是捕获已知插入/删除事件的最强大的仪器。这项研究是当前基因组学技术的基准，也是为实验设计和下一代测序变异调用提供信息的资源。