title: High resolution array-CGH analysis of single cells.
abstract: Heterogeneity in the genome copy number of tissues is of particular importance in solid tumor biology. Furthermore, many clinical applications such as pre-implantation and non-invasive prenatal diagnosis would benefit from the ability to characterize individual single cells. As the amount of DNA from single cells is so small, several PCR protocols have been developed in an attempt to achieve unbiased amplification. Many of these approaches are suitable for subsequent cytogenetic analyses using conventional methodologies such as comparative genomic hybridization (CGH) to metaphase spreads. However, attempts to harness array-CGH for single-cell analysis to provide improved resolution have been disappointing. Here we describe a strategy that combines single-cell amplification using GenomePlex library technology (GenomePlex) Single Cell Whole Genome Amplification Kit, Sigma-Aldrich, UK) and detailed analysis of genomic copy number changes by high-resolution array-CGH. We show that single copy changes as small as 8.3 Mb in single cells are detected reliably with single cells derived from various tumor cell lines as well as patients presenting with trisomy 21 and Prader-Willi syndrome. Our results demonstrate the potential of this technology for studies of tumor biology and for clinical diagnostics.
authors.affiliation: The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK. heike@sanger.ac.uk
authors.forename: Heike
authors.initials: H
authors.lastname: Fiegler
authors.forename: Jochen B
authors.initials: JB
authors.lastname: Geigl
authors.forename: Sabine
authors.initials: S
authors.lastname: Langer
authors.forename: Diane
authors.initials: D
authors.lastname: Rigler
authors.forename: Keith
authors.initials: K
authors.lastname: Porter
authors.forename: Kristian
authors.initials: K
authors.lastname: Unger
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Carter
authors.forename: Michael R
authors.initials: MR
authors.lastname: Speicher
mesh_terms: D002292:Carcinoma, Renal Cell / Q000235:genetics; D045744:Cell Line, Tumor; D002869:Chromosome Aberrations; D020732:Cytogenetic Analysis; D004314:Down Syndrome / Q000235:genetics; D005260:Female; D015894:Genome, Human; D023281:Genomics / Q000379:methods; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics; D008297:Male; D009369:Neoplasms / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D011218:Prader-Willi Syndrome / Q000235:genetics; D011296:Prenatal Diagnosis / Q000379:methods*