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Detection of single‐nucleotide mutations including substitutions and deletions by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Author(s) -
Wada Y.,
Yamamoto M.
Publication year - 1997
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(19971015)11:15<1657::aid-rcm83>3.0.co;2-1
Subject(s) - chemistry , mass spectrometry , desorption , ionization , nucleotide , matrix (chemical analysis) , matrix assisted laser desorption/ionization , surface enhanced laser desorption/ionization , time of flight mass spectrometry , chromatography , analytical chemistry (journal) , protein mass spectrometry , ion , electrospray ionization , gene , organic chemistry , biochemistry , adsorption
Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS) was applied to the detection of mutations involving single nucleotides. For detection of a single‐nucleotide deletion, a normally 44 bp region of the L1CAM gene was amplified in a 50 μL solution, and measurement was carried out on the DNA sample after phenol extraction and ethanol precipitation. A molecular mass decrease of 300 Da corresponding to a single nucleotide was identified in the amplified product of patient DNA. For detection of a substitution, an amplified product from a 50 bp region of the human β‐globin gene was cleaved with restriction endonucleases Hae III and Bsp 1286I. Measurement of a mixture of digested fragments, or restriction fragment mass mapping, clearly identified a heterozygous G/C mutation in the molecular ion signals for both sense and antisense single‐stranded DNAs. The results indicate that MALTI‐TOFMS is feasible for genetic diagnosis of point mutations. © 1997 John Wiley & Sons, Ltd.