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首页> 外文期刊>BMC Biotechnology >Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions
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Combining M-FISH and Quantum Dot technology for fast chromosomal assignment of transgenic insertions

机译：结合M-FISH和Quantum Dot技术实现转基因插入的快速染色体分配

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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.
机译：背景技术通过荧光原位杂交（FISH）对转基因插入物进行物理作图是一种可靠且具有成本效益的技术。通常通过并发G带或通过多色FISH方法实现染色体分配，该方法包括一次将感兴趣的转基因序列与一种或多种染色体特异性探针反复共杂交，直到转基因插入的位置被定位为止。确定。结果在这里，我们报告了在小鼠和大鼠模型中在单细胞水平上对转基因插入进行快速染色体分配的技术发展。这包括一个简化的“单一变性混合杂交”程序，该程序结合了通过Multiplex FISH（M-FISH）进行的多色核型分析，一次可同时，明确地鉴定所有染色体，并使用Quantum Dot（QD）偶联物转基因检测。结论尽管通常已经研究过利用QD纳米晶体的独特光学性质（例如光稳定性和亮度）来改善FISH性能，但据我们所知，这是针对目的设计的分子细胞遗传学方案的首次报道，其中QD和标准有机荧光团的结合使用是专门为辅助基因转移技术而量身定制的。

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