title: Use of microarray hybrid capture and next-generation sequencing to identify the anatomy of a transgene.
abstract: Transgenic animals are extensively used to model human disease. Typically, the transgene copy number is estimated, but the exact integration site and configuration of the foreign DNA remains uncharacterized. When transgenes have been closely examined, some unexpected configurations have been found. Here, we describe a method to recover transgene insertion sites and assess structural rearrangements of host and transgene DNA using microarray hybridization and targeted sequence capture. We used information about the transgene insertion site to develop a polymerase chain reaction genotyping assay to distinguish heterozygous from homozygous transgenic animals. Although we worked with a bacterial artificial chromosome transgenic mouse line, this method can be used to analyse the integration site and configuration of any foreign DNA in a sequenced genome.
authors: Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors: Amanda J
authors: AJ
authors: Dubose
authors: Stephen T
authors: ST
authors: Lichtenstein
authors: Narisu
authors: N
authors: Narisu
authors: Lori L
authors: LL
authors: Bonnycastle
authors: Amy J
authors: AJ
authors: Swift
authors: Peter S
authors: PS
authors: Chines
authors: Francis S
authors: FS
authors: Collins
mesh_terms: D000818:Animals; D022202:Chromosomes, Artificial, Bacterial; D060005:Genotyping Techniques; D059014:High-Throughput Nucleotide Sequencing; D051379:Mice; D008822:Mice, Transgenic; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D019076:Transgenes