A mass spectrometry-based multiplex SNP genotyping by utilizing allele specific ligation and strand displacement amplification

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We herein describe a new mass spectrometry-based method for multiplex SNP genotyping by utilizing allele specific ligation and strand displacement amplification (SDA) reaction. In this method, allele-specific ligation is first performed to discriminate base sequence variations at the SNP site within the PCR-amplified target DNA. The primary ligation probe is extended by a universal primer annealing site while the secondary ligation probe has base sequences as an overhang with a nicking enzyme recognition site and complementary mass marker sequence. The ligation probe pairs are ligated by DNA ligase only at specific allele in the target DNA and the resulting ligated product serves as a template to promote the SDA reaction using a universal primer. This process isothermally amplifies short DNA fragments, called mass markers, to be analyzed by mass spectrometry. By varying the sizes of the mass markers, we successfully demonstrated the multiplex SNP genotyping capability of this method by reliably identifying several BRCA mutations in a multiplex manner with mass spectrometry.
Publisher
ELSEVIER ADVANCED TECHNOLOGY
Issue Date
2017-05
Language
English
Article Type
Article
Keywords

SINGLE-NUCLEOTIDE POLYMORPHISMS; ENHANCED RAMAN-SPECTROSCOPY; PROBE AMPLIFICATION; COMPLEX DISEASES; HUMAN GENOME; DNA; BRCA1; MUTATIONS; BIOSENSOR; SURFACE

Citation

BIOSENSORS & BIOELECTRONICS, v.91, pp.122 - 127

ISSN
0956-5663
DOI
10.1016/j.bios.2016.10.065
URI
http://hdl.handle.net/10203/223579
Appears in Collection
CBE-Journal Papers(저널논문)
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