Publication Date:
2012-12-22
Description:
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.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3600412/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3600412/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Zong, Chenghang -- Lu, Sijia -- Chapman, Alec R -- Xie, X Sunney -- HG005097-1/HG/NHGRI NIH HHS/ -- HG005613-01/HG/NHGRI NIH HHS/ -- R01 HG005097/HG/NHGRI NIH HHS/ -- RC2 HG005613/HG/NHGRI NIH HHS/ -- T32 GM008313/GM/NIGMS NIH HHS/ -- New York, N.Y. -- Science. 2012 Dec 21;338(6114):1622-6. doi: 10.1126/science.1229164.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/23258894" target="_blank"〉PubMed〈/a〉
Keywords:
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
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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