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| Item Type: | Article |
|---|---|
| Title: | Single-nucleotide polymorphisms: analysis by mass spectrometry |
| Creators Name: | Sauer, S., Reinhardt, R., Lehrach, H. and Gut, I.G. |
| Abstract: | 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. |
| Keywords: | Alkylation, DNA Primers, Exonucleases, Genotype, Mass Spectrometry, Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry, Microarray Analysis, Oligonucleotides, Polymerase Chain Reaction, Single Nucleotide Polymorphism, Solid Phase Extraction, Transferases |
| Source: | Nature Protocols |
| ISSN: | 1754-2189 |
| Publisher: | Nature Publishing Group |
| Volume: | 1 |
| Number: | 4 |
| Page Range: | 1761-1771 |
| Date: | 22 November 2006 |
| Official Publication: | https://doi.org/10.1038/nprot.2006.257 |
| PubMed: | View item in PubMed |
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