Fabrication of 3D continuous-flow reverse-transcription polymerase chain reaction microdevice integrated with on-chip fluorescence detection for semi-quantitative assessment of gene expression
- Authors
- Quang Nghia Pham; Kieu The Loan Trinh; Nguyen Khoi Song Tran; Park, Tae-Sik; Lee, Nae Yoon
- Issue Date
- 7-Dec-2018
- Publisher
- ROYAL SOC CHEMISTRY
- Citation
- ANALYST, v.143, no.23, pp.5692 - 5701
- Journal Title
- ANALYST
- Volume
- 143
- Number
- 23
- Start Page
- 5692
- End Page
- 5701
- URI
- https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/2955
- DOI
- 10.1039/c8an01739e
- ISSN
- 0003-2654
- Abstract
- We fabricate a three-dimensional (3D) microdevice operated with minimal peripheral accessories, including a portable pump for semi-automated sample delivery and a single heater for temperature control, for performing reverse transcription polymerase chain reaction (RT-PCR) integrated with a downstream fluorescence detection module for semi-quantitative assessment of gene expression. The microdevice was fabricated by wrapping a polytetrafluoroethylene (PTFE) tube around a pre-designed polycarbonate mold to create a seamless microchannel for both the reverse transcription (RT) of RNA and the amplification of complementary DNA. In addition, a silicone tube, which underwent a two-step surface modification mediated by polyethyleneimine and glutaraldehyde coating, was connected at the outlet to capture amplicons downstream of the PTFE tube for on-site fluorescence detection. This fabrication method enabled continuous-flow RT-PCR (CF RT-PCR) using the 3D CF RT-PCR microdevice as a reactor, a single heater for the temperature control of both RT and PCR processes, and a disposable plastic syringe for semi-automated sample delivery. The microdevice was successfully implemented for the identification of the beta-actin gene, a constitutively expressed gene in all cells, and the sphingosine-1-phosphate lyase 1 gene, a potential pharmacological target gene in the diagnosis of cancer, diabetes, and atherosclerosis. This portable integrated microdevice offers a potential approach towards preliminary studies of gene expression and identification of RNA viruses.
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Collections - 바이오나노대학 > 바이오나노학과 > 1. Journal Articles
- 바이오나노대학 > 생명과학과 > 1. Journal Articles
- 산업·환경대학원 > 산업환경공학과 > 1. Journal Articles
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