Simultaneous Genotyping of Indels and SNPs by Mass Spectroscopy

Nucleotide insertion/deletion polymorphisms (indels) in ApoE gene were precisely genotyped using artificial ribonucleases and MALDI-TOF MS. The RNA fragments for MS analysis were prepared by treating RNA specimens with our artificial ribonucleases, which consist of LuCl 3 (molecular scissors) and ol...

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Veröffentlicht in:Journal of the American Society for Mass Spectrometry 2006, Vol.17 (1), p.3-8
Hauptverfasser: Sasayama, Takuro, Kato, Mayu, Aburatani, Hiroyuki, Kuzuya, Akinori, Komiyama, Makoto
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Sprache:eng
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Zusammenfassung:Nucleotide insertion/deletion polymorphisms (indels) in ApoE gene were precisely genotyped using artificial ribonucleases and MALDI-TOF MS. The RNA fragments for MS analysis were prepared by treating RNA specimens with our artificial ribonucleases, which consist of LuCl 3 (molecular scissors) and oligonucleotides bearing two acridine groups (RNA-activator for site-selective scission). RNA scission by Lu(III) ion always occurred at the phosphodiester linkages in front of the two acridines, even when the RNA specimens involved consecutive cytidine sequences of different lengths. Thus, even complicated mixtures of these indel specimens were completely genotyped by using only one acridine-bearing oligonucleotide and by subjecting the reaction mixture to single MS measurement. Moreover, single nucleotide polymorphism (SNP) in the consecutive sequences could be genotyped simultaneously with the indels.
ISSN:1044-0305
1879-1123
DOI:10.1016/j.jasms.2005.08.016