Matrix-induced fragmentation of P3′-N5′ phosphoramidate-containing DNA: high-throughput MALDI-TOF analysis of genomic sequence polymorphisms
AUTOR(ES)
Shchepinov, Mikhail S.
FONTE
Oxford University Press
RESUMO
Chemical and enzymatic approaches were used to produce polynucleotide fragments containing acid-labile internucleotide P3′-N5′ phosphoramidate bonds, either in a surface-bound form or in solution. The primer extension reaction utilizing 5′-amino-5′-deoxynucleoside 5′-triphosphates generates polynucleotides that can be fragmented into short, easy-to-analyze pieces simply by being premixed with the acidic matrices typically used for MALDI-TOF mass spectrometry of nucleic acids. This leads to detection procedures that are simple, robust and easy to automate. Utilizing this approach, a polymorphic site in the human ADRB3 gene was interrogated. Primer extensions with phosphoramidate analogs of dNTPs allowed for unambiguous discrimination of all possible genotypes.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=55906Documentos Relacionados
- High-Throughput MALDI-TOF Discovery of Genomic Sequence Polymorphisms
- RNase T1 mediated base-specific cleavage and MALDI-TOF MS for high-throughput comparative sequence analysis
- Hierarchical high-throughput SNP genotyping of the human Y chromosome using MALDI-TOF mass spectrometry
- Multiplexed discovery of sequence polymorphisms using base-specific cleavage and MALDI-TOF MS
- High-Throughput Method for Detecting Genomic-Deletion Polymorphisms