Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions
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Abstract
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.
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mohammed2011combiningbmc
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| Authors | Mohammed, Yusuf;LV, Bauer David;M, Lipinski Daniel;E, MacLaren Robert;Richard, Wade-Martins;U, Mir Kalim;V, Volpi Emanuela; |
| Journal | bmc biotechnology |
| Year | 2011 |
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| Keywords | Keywords not found |
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