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Single-nucleotide polymorphisms: analysis by mass spectrometry

MPG-Autoren
http://pubman.mpdl.mpg.de/cone/persons/resource/persons50512

Sauer,  Sascha
Nutrigenomics and Gene Regulation (Sascha Sauer), Independent Junior Research Groups (OWL), Max Planck Institute for Molecular Genetics, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons50488

Reinhardt,  Richard
High Throughput Technologies, Max Planck Institute for Molecular Genetics, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons50409

Lehrach,  Hans
Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Volltexte (frei zugänglich)

Sauer.pdf
(beliebiger Volltext), 336KB

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Zitation

Sauer, S., Reinhardt, R., Lehrach, H., & Gut, I. G. (2006). Single-nucleotide polymorphisms: analysis by mass spectrometry. Nature Protocols, 1(4), 1761-1771. doi:10.1038/nprot.2006.257.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-0010-8516-6
Zusammenfassung
Matrix-assisted laser desorption-ionization (MALDI) mass spectrometry has evolved as a powerful method for analyzing nucleic acids. Here we provide protocols for genotyping single-nucleotide polymorphisms (SNPs) by MALDI based on PCR and primer extension to generate allele-specific products. Furthermore, we present three different approaches for sample preparation of primer-extension products before MALDI analysis and discuss their potential areas of application. The first approach, the 'GOOD' assay, is a purification-free procedure that uses DNA-modification chemistry, including alkylation of phosphorothioate linkages in the extension primers. The other two approaches use either solid-phase extraction or microarray purification for the purification of primer-extension products. Depending on the reaction steps of the various approaches, the protocols take about 6–8 hours.