Genome-wide detection of single-nucleotide and copy-number variations of a single human cell

Science. 2012 Dec 21;338(6114):1622-6. doi: 10.1126/science.1229164.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / genetics
  • Cell Line, Tumor
  • Colorectal Neoplasms / genetics*
  • DNA Copy Number Variations*
  • DNA, Neoplasm / genetics*
  • Genome, Human
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Mutation Rate
  • Nucleic Acid Amplification Techniques / methods*
  • Point Mutation*
  • Polymorphism, Single Nucleotide*
  • Sequence Analysis, DNA / methods
  • Single-Cell Analysis*

Substances

  • DNA, Neoplasm