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Automated solid-phase extraction for purification of single nucleotide polymorphism genotyping products prior to matrix-assisted laser desorption/ionisation time-of-flight mass spectrometric analysis

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

Kepper,  Pamela
Max Planck Society;

Smyra,  Anett
Max Planck Society;

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

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

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

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Zitation

Sauer, S., Kepper, P., Smyra, A., Dahl, A., Ferse, F.-T., Lehrach, H., et al. (2004). Automated solid-phase extraction for purification of single nucleotide polymorphism genotyping products prior to matrix-assisted laser desorption/ionisation time-of-flight mass spectrometric analysis. Journal of Chromatography A, 1049(1-2), 9-16. doi:10.1016/j.chroma.2004.07.069.


Zitierlink: https://hdl.handle.net/11858/00-001M-0000-0010-87C8-6
Zusammenfassung
In this article, we describe the application of a novel micro elution solid-phase extraction method for purification of short stretches of DNA products of single nucleotide polymorphisms (SNPs) prior to MALDI mass spectrometry analysis. An important feature of our method is that the purification columns containing a copolymer of divinylbenzene and N-vinylpyrrolidone can be used several times thereby significantly reducing costs. We implemented this DNA purification technology into a fully automated procedure including molecular biology, MALDI sample preparation, automated mass spectrometric analysis and SNP allele calling by software. Due to the facile purification protocol, the methodology shown could furthermore be used for other applications where efficient medium to high-throughput purification of nucleic acids is required.