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	Surface invasive cleavage assay on a maskless light-directed diamond DNA microarray for genome-wide human SNP mapping
	Nie, Bei1,3; Yang, Min2; Fu, Weiling3; Liang, Zhiqing3
	2015
摘要	The surface invasive cleavage assay, because of its innate accuracy and ability for self-signal amplification, provides a potential route for the mapping of hundreds of thousands of human SNP sites. However, its performance on a high density DNA array has not yet been established, due to the unusual "hairpin" probe design on the microarray and the lack of chemical stability of commercially available substrates. Here we present an applicable method to implement a nanocrystalline diamond thin film as an alternative substrate for fabricating an addressable DNA array using maskless light-directed photochemistry, producing the most chemically stable and biocompatible system for genetic analysis and enzymatic reactions. The surface invasive cleavage reaction, followed by degenerated primer ligation and post-rolling circle amplification is consecutively performed on the addressable diamond DNA array, accurately mapping SNP sites from PCR-amplified human genomic target DNA. Furthermore, a specially-designed DNA array containing dual probes in the same pixel is fabricated by following a reverse light-directed DNA synthesis protocol. This essentially enables us to decipher thousands of SNP alleles in a single-pot reaction by the simple addition of enzyme, target and reaction buffers.
DOI	10.1039/c5an00349k
发表期刊	ANALYST
ISSN	0003-2654
卷号	140期号:13页码:4549-4557
通讯作者	Nie, B (reprint author), Chinese Acad Sci, Chongqing Inst Green & Intelligent Technol, 266 Fangzheng Ave, Chongqing 400714, Peoples R China.
收录类别	SCI
WOS记录号	WOS:000356171400027
语种	英语