Hinweis (Dez. 2019): PubLister wird demnächst abgelöst. Publikationen der Technischen Hochschule Wildau werden zukünftig in HISinOne nachgewiesen. In diesem Zusammenhang kann es momentan zu Überarbeitungen und zu temporären Veränderungen Ihrer persönlichen Publikationslisten kommen. Bei weiteren Fragen wenden Sie sich bitte an friederike.borchert [at] th-wildau [dot] de.

Shining a light on LAMP assays— A comparison of LAMP visualization methods including the novel use of berberine

Title:
Shining a light on LAMP assays— A comparison of LAMP visualization methods including the novel use of berberine
Abstract:
The need for simple and effective assays for detecting nucleic acids by isothermal amplification reactions has led to a great variety of end point and real-time monitoring methods. Here we tested direct and indirect methods to visualize the amplification of potato spindle tuber viroid (PSTVd) by loop-mediated isothermal amplification (LAMP) and compared features important for one-pot in-field applications. We compared the performance of magnesium pyrophosphate, hydroxynaphthol blue (HNB), calcein, SYBR Green I, EvaGreen, and berberine. All assays could be used to distinguish between positive and negative samples in visible or UV light. Precipitation of magnesium-pyrophosphate resulted in a turbid reaction solution. The use of HNB resulted in a color change from violet to blue, whereas calcein induced a change from orange to yellow-green. We also investigated berberine as a nucleic acid-specific dye that emits a fluorescence signal under UV light after a positive LAMP reaction. It has a comparable sensitivity to SYBR Green I and EvaGreen. Based on our results, an optimal detection method can be chosen easily for isothermal real-time or end point screening applications.
Year:
2015
Publication type:
Journal article
Journal:
Biotechniques
ISSN:
0736-6205
Number:
4
Volume:
58
Pages:
189-194
Language:
English
Document status:
Open access
PubListerURL:
https://publister.bib.th-wildau.de/publister/public/publication/873
Keywords:
bioelectrocatalysis; fructose dehydrogenase; sulfonated polyanilines; ITO electrodes; entrapped enzyme
Fischbach, J., Xander, N., Frohme, M., & Glökler, J. (2015). Shining a light on LAMP assays— A comparison of LAMP visualization methods including the novel use of berberine Biotechniques. 58 (4), 189-194.
@article{873,
    author           = {Fischbach, Jens and Xander, Nina Carolin and Frohme, Marcus and Glökler, Jörn Felix},
    title            = {Shining a light on LAMP assays— A comparison of LAMP visualization methods including the novel use of berberine},
    journal          = {Biotechniques },
    year             = {2015},
    volume           = {58},
    number           = {4},
    pages            = {189-194},
    publisher        = {Future Science},
    url              = {http://www.biotechniques.com/BiotechniquesJournal/2015/April/Shining-a-light-on-LAMP-assays--A-comparison-of-LAMP-visualization-methods-including-the-novel-use-of-berberine/biotechniques-357796.html},
}


Search this title in Google Scholar