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A simple and accurate SNP scoring strategy based on typeIIS restriction endonuclease cleavage and matrix-assisted laser desorption/ionization mass spectrometry

Cited 5 time in Web of Science Cited 5 time in Scopus
Authors
Hong, Sun Pyo; Ji, Seung Il; Rhee, Hwanseok; Shin, Soo Kyeong; Hwang, Sun Young; Lee, Seung Hwan; Lee, Soong Deok; Oh, Heung-Bum; Yoo, Wangdon; Kim, Soo-Ok
Issue Date
2008-06-10
Publisher
BioMed Central
Citation
BMC Genomics. 2008 Jun 9;9:276.
Keywords
AllelesBase SequenceDNA/geneticsDNA Primers/geneticsDeoxyribonucleases, Type II Site-SpecificGene Frequency*Genetic TechniquesHaplotypesHumansKoreaMethylenetetrahydrofolate Reductase (NADPH2)/geneticsPolymerase Chain ReactionPolymorphism, Restriction Fragment Length*Polymorphism, Single NucleotideSpectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
Abstract
BACKGROUND: We describe the development of a novel matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF)-based single nucleotide polymorphism (SNP) scoring strategy, termed Restriction Fragment Mass Polymorphism (RFMP) that is suitable for genotyping variations in a simple, accurate, and high-throughput manner. The assay is based on polymerase chain reaction (PCR) amplification and mass measurement of oligonucleotides containing a polymorphic base, to which a typeIIS restriction endonuclease recognition was introduced by PCR amplification. Enzymatic cleavage of the products leads to excision of oligonucleotide fragments representing base variation of the polymorphic site whose masses were determined by MALDI-TOF MS. RESULTS: The assay represents an improvement over previous methods because it relies on the direct mass determination of PCR products rather than on an indirect analysis, where a base-extended or fluorescent report tag is interpreted. The RFMP strategy is simple and straightforward, requiring one restriction digestion reaction following target amplification in a single vessel. With this technology, genotypes are generated with a high call rate (99.6%) and high accuracy (99.8%) as determined by independent sequencing. CONCLUSION: The simplicity, accuracy and amenability to high-throughput screening analysis should make the RFMP assay suitable for large-scale genotype association study as well as clinical genotyping in laboratories.
ISSN
1471-2164 (Electronic)
Language
English
URI
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18538037

http://www.biomedcentral.com/content/pdf/1471-2164-9-276.pdf

http://hdl.handle.net/10371/46312
DOI
https://doi.org/10.1186/1471-2164-9-276
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College of Medicine/School of Medicine (의과대학/대학원)Forensic Medicine (법의학전공)Journal Papers (저널논문_법의학전공)
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