Camille A. Daniels, Adetola A. Abdulkadir, Megan H. Cleveland, Jennifer H. McDaniel, David Jáspez, Luis Alberto Rubio-Rodríguez, Adrián Muñoz-Barrera, José Miguel Lorenzo-Salazar, Carlos Flores, Byunggil Yoo, Sayed Mohammad Ebrahim Sahraeian, Yina Wang, Massimiliano Rossi, Arun Visvanath, Lisa Murray, Wei-Ting Chen, Severine Catreux, James Han, Rami Mehio, Gavin Parnaby, Andrew Carroll, Pi-Chuan Chang, Kishwar Shafin, Daniel Cook, Alexey Kolesnikov, Lucas Brambrink, Mohammed Faizal Eeman Mootor, Yash Patel, Takafumi N. Yamaguchi, Paul C. Boutros, Karolina Sienkiewicz, Jonathan Foox, Christopher E. Mason, Bryan R. Lajoie, Carlos A. Ruiz-Perez, Semyon Kruglyak, Justin M. Zook, Nathan D. Olson
We developed a benchmark set of subclonal variants in the Genome in a Bottle (GIAB) Consortium HG002 reference material (RM) DNA for evaluating lower-frequency variant callsets. We used a somatic variant caller with high-coverage (300×) whole-genome sequencing data from the GIAB Ashkenazi Jewish trio to identify potential subclonal variants in the HG002 RM DNA. Using orthogonal sequencing data and manual curation, we defined a benchmark set with 85 high-confidence subclonal single-nucleotide variants (SNVs) (allele frequency [AF] > 5%) and a benchmark region covering 2.45 Gbp of the autosomes. External validation supported that it can be used to reliably identify both false negatives and false positives for a variety of sequencing technologies and variant callers. By adding our characterization of mosaic SNVs in this widely used cell line, we have expanded the scope of bioinformatic and sequencing applications for which the HG002 GIAB RM can be used to include benchmarking subclonal SNVs.