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Genotyping of Apolipoprotein E by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Author(s) -
Srinivasan Jannavi R.,
Kachman Maureen T.,
Killeen Anthony A.,
Akel Nahida,
Siemieniak David,
Lubman David M.
Publication year - 1998
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/(sici)1097-0231(19980831)12:16<1045::aid-rcm281>3.0.co;2-y
Subject(s) - chemistry , mass spectrometry , surface enhanced laser desorption/ionization , time of flight mass spectrometry , chromatography , desorption , matrix assisted laser desorption/ionization , matrix (chemical analysis) , genotyping , ionization , analytical chemistry (journal) , protein mass spectrometry , tandem mass spectrometry , ion , genotype , biochemistry , adsorption , organic chemistry , gene
The genotyping of the various isoforms of Apolipoprotein E (apo E) has been performed using matrix‐assisted laser desorption/ionization (MALDI‐MS). The polymerase chain reaction was used to amplify the specific apo E gene sequence followed by digestion with Cfo I ( Clostridium formicoaceticum ), for generating restriction fragments for rapid and accurate mass analysis. An exonuclease I digestion step was introduced to remove the unused primers after PCR, which can otherwise interfere in the mass spectral analysis. By replacing the gel electrophoresis detection step with MALDI‐MS, restriction isotyping of the apo E gene was achieved. Genotyping of an unknown sample obtained from an independent diagnostic laboratory demonstrated the validity of the MALDI‐MS method for the routine clinical analysis of apo E. © 1998 John Wiley & Sons, Ltd.