Mutation detection using mass spectrometric separation of tiny oligonucleotide fragments
Details
Publication Year 2002-09, Volume 12, Issue #9, Page 1428-1433
Journal Title
GENOME RESEARCH
Publication Type
Journal Article
Abstract
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 oil 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 call 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.
Publisher
COLD SPRING HARBOR LAB PRESS
Keywords
DNA; POLYMORPHISMS
Publisher's Version
https://doi.org/10.1101/gr.157802
Rights Notice
Refer to copyright notice on published article.


Creation Date: 2002-09-01 12:00:00
Last Modified: 0001-01-01 12:00:00
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