Computational correction of index switching in multiplexed sequencing libraries

2. Xu, J. et al. Nat. Genet. 49, 377–386 (2017). 3. Huang, F.W. et al. Science 339, 957–959 (2013). 4. Stern, J.L., Theodorescu, D., Vogelstein, B., Papadopoulos, N. & Cech, T.R. Genes Dev. 29, 2219–2224 (2015). specific and universal normalization controls (Supplementary Fig. 3). A targeted allelic ATAC assay is highly desirable because only a small fraction of DNA elements (~1%) both possess informative sequence variants and demonstrate allelic regulation2. Inclusion of an informative SNP between primer-binding sites allows for targeted analysis of allele-specific accessibility by ATAC-PCR, for example, to assess the effect of disease-associated variants on allele-specific accessibility. For well-characterized TERT promoter mutations3,4, ATAC-PCR and Sanger sequencing demonstrated that only the mutant allele conferred TERT promoter chromatin accessibility (Fig. 1c, Supplementary Table 4). We further validated allelic ATAC-PCR on a panel of DNA elements with random monoallelic accessibility in hybrid mouse NPC cells2 (Supplementary Table 5). Allele-specific quantification by ATAC-PCR correlated strongly with the allelic accessibility determined by ATAC-seq (R = 0.962) (Supplementary Fig. 4), thus demonstrating accurate quantification of targeted and allelespecific accessibility by ATAC-PCR enabled by APT. Further information on experimental design is available in the Supplementary Methods and the Life Sciences Reporting Summary.