Genome-Wide Detection of Single-Nucleotide and Copy-Number Variations of a Single Human Cell
Year: 2012
Type: article
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.
Source: Science
Authors: Chenghang Zong, Sijia Lu, Alec R. Chapman, X. Sunney Xie
Institution: Harvard University
Language: English
Subfield: Cancer Research
Domain: Life Sciences
Open Access status: green
Awards: Molecular Biophysics Training Grant, Single Cell Single Molecule Digital mRNA Profiling with No PCR Amplification, Harvard University - to develop four advanced microscopy assays, which will report on an individual bacterium's metabolic activity, viability, exposure to antibiotics a..., Real-time single-molecule nucleic acid sequencing with fluorogenic nucleotides, G6916253949, G8203064394
OpenAlex ID: W2064977507