Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions

Yusuf, M., Bauer, D.L.V., Lipinski, D.M., MacLaren, R.E., Wade-Martins, R., Mir, K.U. and Volpi, E. 2011. Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions. BMC Biotechnology. 11, pp. 121-131.

TitleCombining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions
AuthorsYusuf, M., Bauer, D.L.V., Lipinski, D.M., MacLaren, R.E., Wade-Martins, R., Mir, K.U. and Volpi, E.
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.

JournalBMC Biotechnology
Journal citation11, pp. 121-131
ISSN1472-6750
Year2011
PublisherBioMed Central
Publisher's versionVolpi_etal_BMCBiotechnology_2011.pdf
Digital Object Identifier (DOI)doi:10.1186/1472-6750-11-121
Web address (URL)http://dx.doi.org/10.1186/1472-6750-11-121
Publication dates
Published2011

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