Assessment for Melting Temperature Measurement of Nucleic Acid by HRM
Joint Authors
Wang, Jing
Pan, Xiaoming
Liang, Xingguo
Source
Journal of Analytical Methods in Chemistry
Issue
Vol. 2016, Issue 2016 (31 Dec. 2016), pp.1-8, 8 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2016-10-19
Country of Publication
Egypt
No. of Pages
8
Main Subjects
Abstract EN
High resolution melting (HRM), with a high sensitivity to distinguish the nucleic acid species with small variations, has been widely applied in the mutation scanning, methylation analysis, and genotyping.
For the aim of extending HRM for the evaluation of thermal stability of nucleic acid secondary structures on sequence dependence, we investigated effects of the dye of EvaGreen, metal ions, and impurities (such as dNTPs) on melting temperature (Tm) measurement by HRM.
The accuracy of HRM was assessed as compared with UV melting method, and little difference between the two methods was found when the DNA Tm was higher than 40°C.
Both insufficiency and excessiveness of EvaGreen were found to give rise to a little bit higher Tm, showing that the proportion of dye should be considered for precise Tm measurement of nucleic acids.
Finally, HRM method was also successfully used to measure Tms of DNA triplex, hairpin, and RNA duplex.
In conclusion, HRM can be applied in the evaluation of thermal stability of nucleic acid (DNA or RNA) or secondary structural elements (even when dNTPs are present).
American Psychological Association (APA)
Wang, Jing& Pan, Xiaoming& Liang, Xingguo. 2016. Assessment for Melting Temperature Measurement of Nucleic Acid by HRM. Journal of Analytical Methods in Chemistry،Vol. 2016, no. 2016, pp.1-8.
https://search.emarefa.net/detail/BIM-1107336
Modern Language Association (MLA)
Wang, Jing…[et al.]. Assessment for Melting Temperature Measurement of Nucleic Acid by HRM. Journal of Analytical Methods in Chemistry No. 2016 (2016), pp.1-8.
https://search.emarefa.net/detail/BIM-1107336
American Medical Association (AMA)
Wang, Jing& Pan, Xiaoming& Liang, Xingguo. Assessment for Melting Temperature Measurement of Nucleic Acid by HRM. Journal of Analytical Methods in Chemistry. 2016. Vol. 2016, no. 2016, pp.1-8.
https://search.emarefa.net/detail/BIM-1107336
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1107336