Ladder-Shape Melting Temperature Isothermal Amplification of Nucleic Acids | Zendy

Deguo Wang | Zendy; Yongzhen Wang | Zendy; Meng Zhang | Zendy; Yongqing Zhang | Zendy; Juntao Sun | Zendy; Chunmei Song | Zendy; Fugang Xiao | Zendy; Yuan Ping | Zendy; Pan Chen | Zendy; Yushan Hu | Zendy; Chaoqun Wang | Zendy; Yanhong Liu | Zendy

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Ladder-Shape Melting Temperature Isothermal Amplification of Nucleic Acids

Author(s) -

Deguo Wang,

Yongzhen Wang,

Meng Zhang,

Yongqing Zhang,

Juntao Sun,

Chunmei Song,

Fugang Xiao,

Yuan Ping,

Pan Chen,

Yushan Hu,

Chaoqun Wang,

Yanhong Liu

Publication year - 2021

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/btn-2020-0173

Subject(s) - loop mediated isothermal amplification , melting temperature , melting curve analysis , nucleic acid , isothermal process , polymerase chain reaction , nested polymerase chain reaction , dna , biology , microbiology and biotechnology , polymerase , chemistry , genetics , materials science , thermodynamics , physics , gene , composite material

A novel method, termed ladder-shape melting temperature isothermal amplification (LMTIA), was developed in this study. As a proof of concept, one pair of primers or two pairs of nested primers and a thermostable DNA polymerase were employed to amplify the internal transcribed spacer of Oryza sativa with the ladder-shape melting temperature curve. Our results demonstrated that the LMTIA assay with nested primers was 50-fold more sensitive than the LAMP assay with the same level of specificity. The LMTIA method has the potential to be used for the prevention and control of emerging epidemics caused by different types of pathogens.

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