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Mutation detection using mass spectrometric separation of tiny oligonucleotide fragments

journal contribution
posted on 2023-05-16, 20:26 authored by Elso, C, Toohey, B, Reid, GE, Poetter, K, Simpson, RJ, Simon James FooteSimon James Foote
A DNA mutation detection protocol able to identify and characterize a previously unknown change in a given sequence in a rapid, efficient, sensitive, and inexpensive manner is required to take advantage of the resources now available to researchers through the genome sequencing projects. We have developed a method based on base-specific cleavage of polymerase chain reaction (PCR) products and then separation of the fragments by matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS), which can meet these criteria. Differences are seen as the presence, absence, or mass change of peaks corresponding to fragments affected by the base difference. This technique is shown through the detection of a polymorphism in the 3′ untranslated region of IL12p40 from a double-stranded PCR product, and the detection of a single nucleotide polymorphism between two mouse strains. The sensitivity of the technique can be increased with the use of postsource decay, which enables differentiation of two fragments of identical mass but different sequence. The level of specificity and the rapid sample analysis time lend this technique to the mass screening of individuals for sequence changes and, in combination with MS sequencing methods, could be used to facilitate rapid resequencing of DNA.

History

Publication title

Genome Research

Volume

12

Issue

9

Pagination

1428-1433

ISSN

1088-9051

Department/School

Menzies Institute for Medical Research

Publisher

Cold Spring Harbor Lab Press

Place of publication

Publications Dept, 500 Sunnyside Blvd, Woodbury, USA, Ny, 11797-2924

Repository Status

  • Restricted

Socio-economic Objectives

Clinical health not elsewhere classified

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