Masking selected sequence variation by incorporating mismatches into melting analysis probes

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Publication Type Journal Article
School or College School of Medicine
Department Pathology
Program ARUP Institute for Clinical and Experimental Pathology
Creator Margraf, Rebecca L.; Mao, Rong; Highsmith, W. Edward; Holtegaard, Leonard; Wittwer, Carl T.
Title Masking selected sequence variation by incorporating mismatches into melting analysis probes
Date 2005-06-28
Description Hybridization probe melting analysis can be complicated by the presence of sequence variations (benign polymorphisms or other mutations) near the targeted mutation. We investigated the use of 'masking' probes to differentiate alleles with similar probe melting temperatures. Selected sequence variation was masked by incorporating deletions, unmatched nucleotides, or universal bases into hybridization probes. Such masking probes create a probe:target mismatch with all possible alleles at the selected polymorphic location. Any allele with additional variation at another site is identified by a lower probe melting temperature than alleles that vary only at the masked position. This technique was applied to RET and HPA6 mutation detection using unlabeled hybridization probes, a saturating dsDNA dye and high-resolution melting analysis.
Type Text
Publisher Associated and Regional University Pathologists (ARUP) Institute for Clinical and Experimental Pathology
Volume 27
Issue 3
First Page 269
Last Page 278
Subject probe hybridization; high-resolution melting analysis; masking; RET
Subject MESH Polymorphism, Genetic; Mutation; DNA Probes; Proto-Oncogene Proteins c-ret
Language eng
Bibliographic Citation Margraf, R.L., Mao, R., & Wittwer, C.T. (2006). Masking selected sequence variation by incorporating mismatches into melting analysis probes. Human Mutation, 27(3), 269-78. DOI: 10.1002/humu.20290
Rights Management This is a preprint of an article published in Human Mutation copyright 2006. Margraf, R.L., Mao, R., Wittwer, C.T. (2006). Masking selected sequence variation by incorporating mismatches into melting analysis probes. Human Mutation, 27(3), 269-78. http://www3.interscience.wiley.com/cgi-bin/jhome/38515
Format Medium application/pdf
Format Extent 300896 bytes
Identifier ir-main,252
ARK ark:/87278/s6bz6qm9
Setname ir_uspace
Date Created 2012-06-13
Date Modified 2021-05-06
ID 706340
Reference URL https://collections.lib.utah.edu/ark:/87278/s6bz6qm9
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