Integrated RNA extraction and RT-PCR for semi-quantitative gene expression studies on a microfluidic device Shaw, Kristy J., Hughes, Elizabeth M., Dyer, Charlotte E., Greenman, John and Haswell, Stephen J. 2013, Integrated RNA extraction and RT-PCR for semi-quantitative gene expression studies on a microfluidic device, Laboratory investigation, vol. 93, pp. 961-966, doi: 10.1038/labinvest.2013.76. Attached Files Name Description MIMEType Size Downloads Title Integrated RNA extraction and RT-PCR for semi-quantitative gene expression studies on a microfluidic device Author(s) Shaw, Kristy J. Hughes, Elizabeth M. Dyer, Charlotte E. Greenman, John Haswell, Stephen J. Journal name Laboratory investigation Volume number 93 Start page 961 End page 966 Total pages 6 Publisher United States and Canadian Academy of Pathology Place of publication Baltimore, Md. Publication date 2013 ISSN 0023-6837 1530-0307 Keyword(s) diagnostic metabolism microfluidics point-of-care Summary This paper describes the development of a microfluidic methodology, using RNA extraction and reverse transcription PCR, for investigating expression levels of cytochrome P450 genes. Cytochrome P450 enzymes are involved in the metabolism of xenobiotics, including many commonly prescribed drugs, therefore information on their expression is useful in both pharmaceutical and clinical settings. RNA extraction, from rat liver tissue or primary rat hepatocytes, was performed using a silica-based solid-phase extraction technique. Following elution of the purified RNA, amplification of target sequences for the housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the cytochrome P450 gene CYP1A2, was carried out using a one-step reverse transcription PCR. Once the microfluidic methodology had been optimized, analysis of control and 3-methylcholanthrene-induced primary rat hepatocytes were used to evaluate the system. As expected, GAPDH was consistently expressed, whereas CYP1A2 levels were found to be raised in the drug-treated samples. The proposed system offers an initial platform for development of both rapid throughput analyzers for pharmaceutical drug screening and point-of-care diagnostic tests to aid provision of drug regimens, which can be tailor-made to the individual patient. Language eng DOI 10.1038/labinvest.2013.76 Field of Research 060101 Analytical Biochemistry Socio Economic Objective 920203 Diagnostic Methods HERDC Research category C1.1 Refereed article in a scholarly journal Copyright notice ©2013, Nature Publishing Group Free to Read? Yes Persistent URL http://hdl.handle.net/10536/DRO/DU:30063526 Document type: Journal Article Collections: Office of the Deputy Vice-Chancellor (Research) Open Access Collection Related Links Link Description Link to full-text (open access)   Connect to published version (restricted access) Go to link with your DU access privileges     Unless expressly stated otherwise, the copyright for items in DRO is owned by the author, with all rights reserved. Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au. Versions Version Filter Type Fri, 23 May 2014, 09:38:07 EST Fri, 23 May 2014, 10:56:08 EST Mon, 09 Jun 2014, 13:25:37 EST Mon, 09 Jun 2014, 13:27:04 EST Thu, 11 Dec 2014, 20:21:27 EST Wed, 18 Nov 2015, 07:27:37 EST Sat, 25 Nov 2017, 01:39:15 EST Sat, 09 Nov 2019, 09:36:08 EST Thu, 06 Jan 2022, 20:36:45 EST Sun, 20 Feb 2022, 20:36:34 EST Filtered Full Citation counts:   Cited 13 times in TR Web of Science Cited 12 times in Scopus Search Google Scholar Access Statistics: 244 Abstract Views, 2 File Downloads  -  Detailed Statistics Created: Fri, 23 May 2014, 09:38:07 EST