Journal Article

The GIAB genomic stratifications resource for human reference genomes

Sedlazeck, Fritz J., Jadhav, Bharati, Huang, Wenyu (Eddy), Dwarshuis, Nathan, Zook, Justin M., McDaniel, Jennifer, Sanio, Philippe, Busby, Ben, Majidian, Sina, Kalra, Divya, Alvarez Jerez, Pilar, Olson, Nathan D., Wagner, Justin, Mondal, Rajarshi

2024

Overview

Despite the growing variety of sequencing and variant-calling tools, no workflow performs equally well across the entire human genome. Understanding context-dependent performance is critical for enabling researchers, clinicians, and developers to make informed tradeoffs when selecting sequencing hardware and software. Here we describe a set of “stratifications,” which are BED files that define distinct contexts throughout the genome. We define these for GRCh37/38 as well as the new T2T-CHM13 reference, adding many new hard-to-sequence regions which are critical for understanding performance as the field progresses. Specifically, we highlight the increase in hard-to-map and GC-rich stratifications in CHM13 relative to the previous references. We then compare the benchmarking performance with each reference and show the performance penalty brought about by these additional difficult regions in CHM13. Additionally, we demonstrate how the stratifications can track context-specific improvements over different platform iterations, using Oxford Nanopore Technologies as an example. The means to generate these stratifications are available as a snakemake pipeline at https://github.com/usnistgov/giab-stratifications . We anticipate this being useful in enabling precise risk-reward calculations when building sequencing pipelines for any of the commonly-used reference genomes. The GIAB genomic stratification resource defines challenging regions in three commonly used human genome references, including the first complete human genome (CHM13). These help understand strengths and weaknesses of sequencing and analysis methods.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 45 / 49 / 631/114/129 / 631/114/2416 / 631/1647/2217 / 631/208/726 / 631/61/212 / Benchmarking / Context / Gene sequencing / Genome, Human / Genomes / Genomic analysis / Genomics - methods / High-Throughput Nucleotide Sequencing - methods / Human performance / Humanities and Social Sciences / Humans / multidisciplinary / Nucleotide sequence / Science / Science (multidisciplinary) / Sequence Analysis, DNA - methods / Software / Workflow