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Detection of simultaneous multi-mutations using base-quenched probe

Mao, Huihui ; Luo, Guanghua ; Zhang, Jun and Xu, Ning LU (2018) In Analytical Biochemistry 543. p.79-81
Abstract

The base-quenched probe method for detecting single nucleotide polymorphisms (SNPs) relies on real-time PCR and melting-curve approaches. Here, we applied the most common commercial fluorophores including FAM, HEX, CY5, CY3, TET, JOE, Texas Red and ROX for labeling probes to detect multi-mutations simultaneously according to the different fluorescence channels. Accuracy of the method was confirmed by direct sequencing. The results demonstrated that all above dyes could be influenced by bases and could be applied to detect SNPs. Furthermore, this method was applied to detect APOM rs707921, APOM rs707922 and MCP-1 rs1024611 simultaneously, which was demonstrated successfully.

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author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Melting-curve analysis, Single nucleotide polymorphism, The base-quenched probe
in
Analytical Biochemistry
volume
543
pages
3 pages
publisher
Elsevier
external identifiers
  • pmid:29233678
  • scopus:85037820456
ISSN
0003-2697
DOI
10.1016/j.ab.2017.12.011
language
English
LU publication?
yes
id
4b93c4bd-aab5-4908-af20-099312fd658d
date added to LUP
2018-01-02 13:54:56
date last changed
2024-10-14 17:57:02
@article{4b93c4bd-aab5-4908-af20-099312fd658d,
  abstract     = {{<p>The base-quenched probe method for detecting single nucleotide polymorphisms (SNPs) relies on real-time PCR and melting-curve approaches. Here, we applied the most common commercial fluorophores including FAM, HEX, CY5, CY3, TET, JOE, Texas Red and ROX for labeling probes to detect multi-mutations simultaneously according to the different fluorescence channels. Accuracy of the method was confirmed by direct sequencing. The results demonstrated that all above dyes could be influenced by bases and could be applied to detect SNPs. Furthermore, this method was applied to detect APOM rs707921, APOM rs707922 and MCP-1 rs1024611 simultaneously, which was demonstrated successfully.</p>}},
  author       = {{Mao, Huihui and Luo, Guanghua and Zhang, Jun and Xu, Ning}},
  issn         = {{0003-2697}},
  keywords     = {{Melting-curve analysis; Single nucleotide polymorphism; The base-quenched probe}},
  language     = {{eng}},
  month        = {{02}},
  pages        = {{79--81}},
  publisher    = {{Elsevier}},
  series       = {{Analytical Biochemistry}},
  title        = {{Detection of simultaneous multi-mutations using base-quenched probe}},
  url          = {{http://dx.doi.org/10.1016/j.ab.2017.12.011}},
  doi          = {{10.1016/j.ab.2017.12.011}},
  volume       = {{543}},
  year         = {{2018}},
}