A New Workflow for Whole-Genome Sequencing of Single Human Cells

Unbiased amplification of the hok-genome o amplification (WGA) of single cells is crucial to study cancer evolution and genetic heterogeneity, but is challenging due to the high complexity of the human genome. Here, we present a new workflow combining an efficient adapter-linker PCR-based \VGA method with secondgeneration sequencing. This approach allows comparison of single cells at base pair resolution. Amplification recovered up to 74% of the human genome. Copy-number variants and loss of heterozygosity detected in single cell genomes showed concordance of up to 99% to pooled ge nornic DNA. Allele frequencies of mutations could be determined accurately due to an allele dropout rate of only 2%, clearly demonstrating the low bias of our PC11, based AVGA approach. Sequencing with paired-end reads allowed genome-wide analysis of structural variants. By direct comparison to other 1VGA methods, we further endorse its suitability to analyze genetic heterogeneity. (C) 2014 wiley periodicals, Inc.