An Integrated Microdevice for Nucleic Acid Extraction and Amplification from Harmful Algae Cochlodinium polykrikoides

Abstract

Red tides, or harmful algal blooms (HABs), have been known as the notorious term for describing the bursting occurrence of Cochlodinium polykrikoides &#8211 a toxic dinoflagellate species in the SouthSea of Korea since 1995. These algal blooms cause fish kill mainly during late spring and summerand become more widespread, resulting in more damages to fisheries. In this study, a monolithicthermoplastic microdevice was designed to handle both DNA extraction and polymerase chainreaction (PCR) processes in one continuous flow. C. polykrikoides cells were first captured onamine-functionalized microbeads and then lysed to release DNA into subsequent PCR channel. Inthis experiment, Escherichia coli bacterium was used as a model system to illustrate cell adsorption onto the amine-functionalized microbeads because both the E. coli and the algal cell possess negative charges on their surfaces. 244-bp and 216-bp DNA fragments were successfully amplified from C. polykrikoides on a polycarbonate (PC) microdevice following the on-chip extraction, and two processes &#8211 DNA extraction and amplification &#8211 were realized in one continuous flow using the microdevice.blooms cause fish kill mainly during late spring and summerand become more widespread, resulting in more damages to fisheries. In this study, a monolithicthermoplastic microdevice was designed to handle both DNA extraction and polymerase chainreaction (PCR) processes in one continuous flow. C. polykrikoides cells were first captured onamine-functionalized microbeads and then lysed to release DNA into subsequent PCR channel. Inthis experiment, Escherichia coli bacterium was used as a model system to illustrate cell adsorption onto the amine-functionalized microbeads because both the E. coli and the algal cell possess negative charges on their surfaces. 244-bp and 216-bp DNA fragments were successfully amplified from C. polykrikoides on a polycarbonate (PC) microdevice following the on-chip extraction, and two processes &#8211 DNA extraction and amplification &#8211 were realized in one continuous flow using the microdevice.