|Title||Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions|
|Authors||Yusuf, M., Bauer, D.L.V., Lipinski, D.M., MacLaren, R.E., Wade-Martins, R., Mir, K.U. and Volpi, E.|
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.
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.
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.
|Journal citation||11, pp. 121-131|
|Digital Object Identifier (DOI)||doi:10.1186/1472-6750-11-121|
|Web address (URL)||http://dx.doi.org/10.1186/1472-6750-11-121|