Detection of simultaneous multi-mutations using base-quenched probe
(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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https://lup.lub.lu.se/record/4b93c4bd-aab5-4908-af20-099312fd658d
- author
- Mao, Huihui ; Luo, Guanghua ; Zhang, Jun and Xu, Ning LU
- organization
- publishing date
- 2018-02-15
- 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
-
- scopus:85037820456
- pmid:29233678
- 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}}, }