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High specific genotyping method using short target probe and helper probe

Title
High specific genotyping method using short target probe and helper probe
Author
황승용
Keywords
Genotyping; Fluorescence melting curve analysis; DNA melting temperature; Real-time polymerase chain reaction; Fluorescence resonance energy transfer
Issue Date
2016-08
Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
Citation
MOLECULAR AND CELLULAR PROBES, v. 30, NO. 4, Page. 273-276
Abstract
Differentiating 1-bp differences using real-time PCR often leads to false-positive results. Therefore, we developed a fluorescence melting curve analysis (FMCA) method with a short target probe and helper probe labeled with a fluorophore and quencher, respectively. This fluorophore and quencher were designed to be near each other when the probes were hybridized to template DNA. The target probe was designed with a shorter length to facilitate a dramatic shift in melting temperature (Tm) upon encountering mismatched hybridization. In FMCA, when the temperature approached the target probe Tm, the target probe would begin to denature from the template DNA, and at the target probe Tm, the fluorescence signal increased markedly. Here, we examined 1-bp differences using the developed method with mitochondrial DNA from Larimichthys polyactis and Larimichthys crocea. Application of this method permitted specific genotype identification for all cases with no cross-reactivity, even when both templates were added to the same tube. (C) 2016 Elsevier Ltd.
URI
https://www.sciencedirect.com/science/article/pii/S0890850816300305https://repository.hanyang.ac.kr/handle/20.500.11754/75121
ISSN
0890-8508
DOI
10.1016/j.mcp.2016.05.002
Appears in Collections:
GRADUATE SCHOOL[S](대학원) > BIONANOTECHNOLOGY(바이오나노학과) > Articles
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