Fabrication of 3D continuous-flow reverse-transcription polymerase chain reaction microdevice integrated with on-chip fluorescence detection for semi-quantitative assessment of gene expression
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Quang Nghia Pham | - |
dc.contributor.author | Kieu The Loan Trinh | - |
dc.contributor.author | Nguyen Khoi Song Tran | - |
dc.contributor.author | Park, Tae-Sik | - |
dc.contributor.author | Lee, Nae Yoon | - |
dc.date.available | 2020-02-27T08:40:55Z | - |
dc.date.created | 2020-02-06 | - |
dc.date.issued | 2018-12-07 | - |
dc.identifier.issn | 0003-2654 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/2955 | - |
dc.description.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. | - |
dc.language | 영어 | - |
dc.language.iso | en | - |
dc.publisher | ROYAL SOC CHEMISTRY | - |
dc.relation.isPartOf | ANALYST | - |
dc.subject | RT-PCR | - |
dc.subject | A-CHIP | - |
dc.subject | RNA AMPLIFICATION | - |
dc.subject | MICROCHANNEL | - |
dc.subject | LIQUID | - |
dc.subject | DEVICE | - |
dc.subject | EXTRACTION | - |
dc.subject | SYSTEMS | - |
dc.subject | VIRUSES | - |
dc.subject | SYRINGE | - |
dc.title | Fabrication of 3D continuous-flow reverse-transcription polymerase chain reaction microdevice integrated with on-chip fluorescence detection for semi-quantitative assessment of gene expression | - |
dc.type | Article | - |
dc.type.rims | ART | - |
dc.description.journalClass | 1 | - |
dc.identifier.wosid | 000450658000027 | - |
dc.identifier.doi | 10.1039/c8an01739e | - |
dc.identifier.bibliographicCitation | ANALYST, v.143, no.23, pp.5692 - 5701 | - |
dc.identifier.scopusid | 2-s2.0-85056573903 | - |
dc.citation.endPage | 5701 | - |
dc.citation.startPage | 5692 | - |
dc.citation.title | ANALYST | - |
dc.citation.volume | 143 | - |
dc.citation.number | 23 | - |
dc.contributor.affiliatedAuthor | Quang Nghia Pham | - |
dc.contributor.affiliatedAuthor | Kieu The Loan Trinh | - |
dc.contributor.affiliatedAuthor | Nguyen Khoi Song Tran | - |
dc.contributor.affiliatedAuthor | Park, Tae-Sik | - |
dc.contributor.affiliatedAuthor | Lee, Nae Yoon | - |
dc.type.docType | Article | - |
dc.subject.keywordPlus | RT-PCR | - |
dc.subject.keywordPlus | A-CHIP | - |
dc.subject.keywordPlus | RNA AMPLIFICATION | - |
dc.subject.keywordPlus | MICROCHANNEL | - |
dc.subject.keywordPlus | LIQUID | - |
dc.subject.keywordPlus | DEVICE | - |
dc.subject.keywordPlus | EXTRACTION | - |
dc.subject.keywordPlus | SYSTEMS | - |
dc.subject.keywordPlus | VIRUSES | - |
dc.subject.keywordPlus | SYRINGE | - |
dc.relation.journalResearchArea | Chemistry | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Analytical | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.
1342, Seongnam-daero, Sujeong-gu, Seongnam-si, Gyeonggi-do, Republic of Korea(13120)031-750-5114
COPYRIGHT 2020 Gachon University All Rights Reserved.
Certain data included herein are derived from the © Web of Science of Clarivate Analytics. All rights reserved.
You may not copy or re-distribute this material in whole or in part without the prior written consent of Clarivate Analytics.