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Development of simple and efficient Lab-on-a-Disc platforms for automated chemical cell lysis
Cell lysis is the most important first step for molecular biology and diagnostic testing. Recently, microfluidic systems have attracted considerable attention due to advantages associated with automation, integration and miniaturization, especially in resource-limited settings. In this work, novel c...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7338454/ https://www.ncbi.nlm.nih.gov/pubmed/32632169 http://dx.doi.org/10.1038/s41598-020-67995-3 |
Sumario: | Cell lysis is the most important first step for molecular biology and diagnostic testing. Recently, microfluidic systems have attracted considerable attention due to advantages associated with automation, integration and miniaturization, especially in resource-limited settings. In this work, novel centrifugal microfluidic platforms with new configurations for chemical cell lysis are presented. The developed systems employ passive form of pneumatic and inertial forces for effective mixing of lysis reagents and cell samples as well as precise fluidic control. Characterizations of the developed Lab-on-a-Discs (LoaDs) have been conducted with dyed deionized (DI) waters and white blood cells (WBCs) to demonstrate the suitability of the proposed systems in terms of mixing, fluidic control and chemical cell lysis. By making comparison between the results of a well-established manual protocol for chemical cell lysis and the proposed chemical cell lysis discs, it has been proved that the developed systems are capable of realizing automated cell lysis with high throughput in terms of proper values of average DNA yield (ranging from 20.6 to 29.8 ng/µl) and purity (ranging from 1.873 to 1.907) as well as suitability of the released DNA for polymerase chain reaction (PCR). By considering the manual chemical lysis protocol as a reference, the efficiency of the LoaDs has been determined 95.5% and 91% for 10 min and 5 min lysis time, respectively. The developed LoaDs provide simple, efficient, and fully automated chemical cell lysis units, which can be easily integrated into operational on-disc elements to obtain sample-to answer settings systems. |
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