Nucleic Acids Research Advance Access published online on August 18, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn518
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Methods Online |
Studying copy number variations using a nanofluidic platform
Fluidigm Corporation, South San Francisco, CA 94080, USA
*To whom correspondence should be addressed. Tel: +1 650 266 6084; Fax: +1 650 871 7152; Email: ramesh.ramakrishnan{at}fluidigm.com
Received June 27, 2008. Revised July 28, 2008. Accepted July 29, 2008.
Copy number variations (CNVs) in the human genome are conventionally detected using high-throughput scanning technologies, such as comparative genomic hybridization and high-density single nucleotide polymorphism (SNP) microarrays, or relatively low-throughput techniques, such as quantitative polymerase chain reaction (PCR). All these approaches are limited in resolution and can at best distinguish a twofold (or 50%) difference in copy number. We have developed a new technology to study copy numbers using a platform known as the digital array, a nanofluidic biochip capable of accurately quantitating genes of interest in DNA samples. We have evaluated the digital array's performance using a model system, to show that this technology is exquisitely sensitive, capable of differentiating as little as a 15% difference in gene copy number (or between 6 and 7 copies of a target gene). We have also analyzed commercial DNA samples for their CYP2D6 copy numbers and confirmed that our results were consistent with those obtained independently using conventional techniques. In a screening experiment with breast cancer and normal DNA samples, the ERBB2 gene was found to be amplified in about 35% of breast cancer samples. The use of the digital array enables accurate measurement of gene copy numbers and is of significant value in CNV studies.