MALDI mass spectrometry analysis of single nucleotide polymorphisms by photocleavage and charge-tagging

AUTOR(ES)

Sauer, Sascha

FONTE

Oxford University Press

RESUMO

High-throughput procedures are an important requirement for future large-scale genetic studies such as genotyping of single nucleotide polymorphisms (SNPs). Matrix-assisted laser desorption/ ionisation mass spectrometry (MALDI-MS) has revolutionised the analysis of biomolecules and, in particular, provides a very attractive solution for the rapid typing of DNA. The analysis of DNA by MALDI can be significantly facilitated by a procedure termed ‘charge-tagging’. We show here a novel approach for the generation of charge-tagged DNA using a photocleavable linker and its implementation in a molecular biological procedure for SNP genotyping consisting of PCR, primer extension, photocleavage and a chemical reaction prior to MALDI target preparation and analysis. The reaction sequence is amenable to liquid handling automation and requires no stringent purification procedures. We demonstrate this new method on SNPs in two genes involved in complex traits.

Documentos Relacionados

• MALDI-TOF mass spectrometric typing of single nucleotide polymorphisms with mass-tagged ddNTPs.
• Facile method for automated genotyping of single nucleotide polymorphisms by mass spectrometry
• DNA sequence analysis by MALDI mass spectrometry.
• Re-sequencing of multiple single nucleotide polymorphisms by liquid chromatography–electrospray ionization mass spectrometry
• Analysis and accurate quantification of CpG methylation by MALDI mass spectrometry