1. Techniques for Evaluation of LAMP Amplicons and their Applications in Molecular Biology
- Author
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Majid Sadeghizadeh, Narges Bodaghabadi, Motalebzadeh H, Dehghan Esmatabadi Mj, Ali Bozorgmehr, Sadegh Babashah, and Baharak Farhangi
- Subjects
Electrophoresis, Agar Gel ,Cancer Research ,Staining and Labeling ,Epidemiology ,Public Health, Environmental and Occupational Health ,Loop-mediated isothermal amplification ,Enzyme-Linked Immunosorbent Assay ,DNA ,Nucleic acid amplification technique ,Amplicon ,Biology ,Fluorescence ,Molecular biology ,Electrophoresis ,Förster resonance energy transfer ,Oncology ,Agarose gel electrophoresis ,Fluorescence Resonance Energy Transfer ,Humans ,Nucleic Acid Amplification Techniques ,Polyacrylamide gel electrophoresis ,DNA Primers - Abstract
Loop-mediated isothermal amplification (LAMP) developed by Notomi et al. (2000) has made it possible to amplify DNA with high specificity, efficiency and rapidity under isothermal conditions. The ultimate products of LAMP are stem-loop structures with several inverted repeats of the target sequence and cauliflower-like patterns with multiple loops shaped by annealing between every other inverted repeats of the amplified target in the similar strand. Because the amplification process in LAMP is achieved by using four to six distinct primers, it is expected to amplify the target region with high selectivity. However, evaluation of reaction accuracy or quantitative inspection make it necessary to append other procedures to scrutinize the amplified products. Hitherto, various techniques such as turbidity assessment in the reaction vessel, post-reaction agarose gel electrophoresis, use of intercalating fluorescent dyes, real-time turbidimetry, addition of cationic polymers to the reaction mixture, polyacrylamide gel-based microchambers, lateral flow dipsticks, fluorescence resonance energy transfer (FRET), enzyme-linked immunosorbent assays and nanoparticle-based colorimetric tests have been utilized for this purpose. In this paper, we reviewed the best-known techniques for evaluation of LAMP amplicons and their applications in molecular biology beside their advantages and deficiencies. Regarding the properties of each technique, the development of innovative prompt, cost-effective and precise molecular detection methods for application in the broad field of cancer research may be feasible.
- Published
- 2015