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Development towards label- and amplification-free genotyping of genomic DNA

机译:向基因组DNA的无标记和无扩增基因分型发展

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Peptide nucleic acid (PNA) microarrays have been used for the analysis of unlabelled DNA molecules by time-of-flight secondary mass spectrometry (ToF-SIMS). PNAs were synthesised with an automated system in multi-well plates followed by the spotting full-length molecules only onto derivatised silicon surfaces using a common split-pin contact printing instrument. After DNA hybridisation, phosphate specific signals were detected and visualised by ToF-SIMS imaging. Because phosphorus is entirely missing in PNAs but is an integral part of nucleic acids, specific signals were only detected on spots at which DNA bound to complementary PNA probes. Combining PNA microarrays with ToF-SIMS detection enables the sensitive analysis of DNA or RNA targets without the need for introducing labels like fluorescent groups. (c) 2006 Elsevier B.V. All rights reserved.
机译:肽核酸(PNA)微阵列已用于通过飞行时间二次质谱(ToF-SIMS)分析未标记的DNA分子。使用自动化系统在多孔板上合成PNA,然后使用普通的分针式接触印刷仪将全长分子仅点到衍生化的硅表面上。 DNA杂交后,检测磷酸盐特异性信号并通过ToF-SIMS成像将其可视化。由于磷在PNA中完全缺失,但却是核酸的组成部分,因此仅在与互补PNA探针结合的DNA上检测到特异性信号。将PNA微阵列与ToF-SIMS检测结合使用,可以对DNA或RNA目标进行灵敏的分析,而无需引入荧光基团等标记。 (c)2006 Elsevier B.V.保留所有权利。

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