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 |
ID |
704112 |
Reference URL |
https://collections.lib.utah.edu/ark:/87278/s6320d5b |