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Info. Vol.14 - No.2 (2020.06.20)
Title Separation, Purification, and Detection of cfDNA in a Microfluidic Device
Authors Hyeon Gi Kye , Christian D. Ahrberg, Byeong Seon Park, Jong Min Lee & Bong Geun Chung*
Institutions Department of Mechanical Engineering, Sogang University, 35 Baekbeom-ro, Mapo-gu, Seoul, 04107, Korea
These authors contributed equally.
*Correspondence and requests for materials should be addressed to B.G. Chung (bchung@sogang.ac.kr)
Abstract Cell free DNA (cfDNA) is degraded DNA fragments found in the blood plasma of cancer patients. While cfDNA is a good marker for early diagnostics and cancer prognosis, the extraction of cfDNA from whole blood and sample preparation for later sequencing is still challenging. Here, we presented a microfluidic device for the removal of cells from a cfDNA sample in a first step. In a second step, carboxylated magnetic beads were used for cfDNA extraction and purification. Lastly, cfDNA was amplified using a lowpower, plasmonic polymerase chain reaction (PCR) system. Using fluorescent-labeled beads, we demonstrated that the separation efficiency for cells was 99% and the mixing efficiency for purification of cfDNA was 94%. Captured cfDNA could be successfully amplified by PCR, as demonstrated by gel electrophoresis. We confirmed that the limit of detection of our microfluidic system was 10 ng/mL.
Keyword Separation, Purification, cfDNA, PCR, Microfluidic device
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