Genome-Wide Detection of Single-Nucleotide and Copy-Number Variations of a Single Human Cell
Menée sur une lignée cellulaire, cette étude évalue la faisabilité d'une technique d'amplification appelée MALBAC pour détecter des variations de nombres de copies de gènes et de simples nucléotides à l'échelle d'une seule cellule
Kindred cells can have different genomes because of dynamic changes in DNA. Single-cell sequencing is needed to characterize these genomic differences but has been hindered by whole-genome amplification bias, resulting in low genome coverage. Here, we report on a new amplification method—multiple annealing and looping-based amplification cycles (MALBAC)—that offers high uniformity across the genome. Sequencing MALBAC-amplified DNA achieves 93% genome coverage ≥1x for a single human cell at 25x mean sequencing depth. We detected digitized copy-number variations (CNVs) of a single cancer cell. By sequencing three kindred cells, we were able to identify individual single-nucleotide variations (SNVs), with no false positives detected. We directly measured the genome-wide mutation rate of a cancer cell line and found that purine-pyrimidine exchanges occurred unusually frequently among the newly acquired SNVs.
Science , résumé, 2012