eCite Digital Repository

Mutation detection using mass spectrometric separation of tiny oligonucleotide fragments


Elso, C and Toohey, B and Reid, GE and Poetter, K and Simpson, RJ and Foote, SJ, Mutation detection using mass spectrometric separation of tiny oligonucleotide fragments, Genome Research, 12, (9) pp. 1428-1433. ISSN 1088-9051 (2002) [Refereed Article]

DOI: doi:10.1101/gr.GR-1578R


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.

Item Details

Item Type:Refereed Article
Research Division:Biomedical and Clinical Sciences
Research Group:Clinical sciences
Research Field:Medical genetics (excl. cancer genetics)
Objective Division:Health
Objective Group:Clinical health
Objective Field:Clinical health not elsewhere classified
UTAS Author:Foote, SJ (Professor Simon Foote)
ID Code:48744
Year Published:2002
Web of Science® Times Cited:10
Deposited By:Menzies Institute for Medical Research
Deposited On:2007-08-01
Last Modified:2011-10-06

Repository Staff Only: item control page