title: Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions.
abstract: BACKGROUND
  Physical mapping of transgenic insertions by Fluorescence in situ Hybridization (FISH) is a reliable and cost-effective technique. Chromosomal assignment is commonly achieved either by concurrent G-banding or by a multi-color FISH approach consisting of iteratively co-hybridizing the transgenic sequence of interest with one or more chromosome-specific probes at a time, until the location of the transgenic insertion is identified.
  
  
  RESULTS
  Here we report a technical development for fast chromosomal assignment of transgenic insertions at the single cell level in mouse and rat models. This comprises a simplified 'single denaturation mixed hybridization' procedure that combines multi-color karyotyping by Multiplex FISH (M-FISH), for simultaneous and unambiguous identification of all chromosomes at once, and the use of a Quantum Dot (QD) conjugate for the transgene detection.
  
  
  CONCLUSIONS
  Although the exploitation of the unique optical properties of QD nanocrystals, such as photo-stability and brightness, to improve FISH performance generally has been previously investigated, to our knowledge this is the first report of a purpose-designed molecular cytogenetic protocol in which the combined use of QDs and standard organic fluorophores is specifically tailored to assist gene transfer technology.
authors.affiliation: Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Yusuf
authors.forename: David L V
authors.initials: DL
authors.lastname: Bauer
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Lipinski
authors.forename: Robert E
authors.initials: RE
authors.lastname: MacLaren
authors.forename: Richard
authors.initials: R
authors.lastname: Wade-Martins
authors.forename: Kalim U
authors.initials: KU
authors.lastname: Mir
authors.forename: Emanuela V
authors.initials: EV
authors.lastname: Volpi
mesh_terms: D000818:Animals; D003116:Color; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D051379:Mice; D016254:Mutagenesis, Insertional / Q000235:genetics*; D020161:Physical Chromosome Mapping / Q000379:methods*; D045663:Quantum Dots; D051381:Rats; D019076:Transgenes / Q000235:genetics*