Masking techniques: masking selected sequence variation by incorporating mismatches into melting analysis probes

Update item information
Publication Type Report
School or College School of Medicine
Department Pathology
Program ARUP Institute for Clinical and Experimental Pathology
Creator Margraf, Rebecca L.; Mao, Rong; Wittwer, Carl T.
Title Masking techniques: masking selected sequence variation by incorporating mismatches into melting analysis probes
Date 2005-10-21
Description Hybridization probe melting analysis can be complicated by the presence of sequence variation (non-pathogenic polymorphisms or other mutations) near the targeted mutation. We investigated the use of 'masking' probes to differentiate alleles with similar probe melting temperatures. Materials and Methods: Selected sequence variation was masked by incorporating deletions, unmatched (non-complementary) 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 proto-oncogene 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
Subject probe hybridization; high-resolution melting analysis; masking; RET
Subject LCSH Chromosome polymorphism; Mutation (Biology); DNA probes; Proto-oncogenes
Subject MESH Polymorphism, Genetic; Mutation; DNA Probes; Proto-Oncogene Proteins c-ret
Language eng
Rights Management http://creativecommons.org/licenses/by-nc-nd/2.5/
Format Medium application/pdf
Identifier ir-main,1
ARK ark:/87278/s6320d5b
Setname ir_uspace
Date Created 2012-06-13
Date Modified 2012-06-13
ID 704112
Reference URL https://collections.lib.utah.edu/ark:/87278/s6320d5b
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