S-Space College of Engineering/Engineering Practice School (공과대학/대학원) Nano-Science and Technology (협동과정 나노과학·기술전공) Journal Papers (저널논문_협동과정 나노과학·기술전공)
Direct electrophoretic microRNA preparation from clinical samples using nanofilter membrane
- Lee, Kidan; Kang, Jae-Hyun; Kim, Hyun-Mi; Ahn, Junhyoung; Lim, Hyungjun; Lee, JaeJong; Jeon, Wan-Jin; Lee, Jae-Hoon; Kim, Ki-Bum
- Issue Date
- Springer Open
- Nano Convergence, 7(1):1
- Nucleic acid extraction; Nucleic acid preparation; miRNA extraction; Nanofltration; Nanoporous membrane; Liquid biopsy
- A method to directly collect negatively charged nucleic acids, such as DNA and RNA, in the biosamples simply by applying an electric field in between the sample and collection buffer separated by the nanofilter membrane is proposed. The nanofilter membrane was made of low-stress silicon nitride with a thickness of 100 nm, and multiple pores were perforated in a highly arranged pattern using nanoimprint technology with a pore size of 200 nm and a pore density of 7.22 × 108/cm2. The electrophoretic transport of hsa-mir-93-5p across the membrane was confirmed in pure microRNA (miRNA) mimic solution using quantitative reverse transcription-polymerase chain reactions (qRT-PCR). Consistency of the collected miRNA quantity, stability of the system during the experiment, and yield and purity of the prepared sample were discussed in detail to validate the effectiveness of the electrical protocol. Finally, in order to check the applicability of this method to clinical samples, liquid biopsy process was demonstrated by evaluating the miRNA levels in sera of hepatocellular carcinoma patients and healthy controls. This efficient system proposed a simple, physical idea in preparation of nucleic acid from biosamples, and demonstrated its compatibility to biological downstream applications such as qRT-PCR as the conventional nucleic acid extraction protocols.