NDLI: Real-time PCR array chip with capillary-driven sample loading and reactor sealing for point-of-care applications

Content Provider	Springer Nature Link
Author	Ramalingam, Naveen Liu, Hao Bing Dai, Chang Chun Jiang, Yu Wang, Hui Wang, Qinghui M Hui, Kam Gong, Hai Qing
Copyright Year	2009
Abstract	A major challenge for the lab-on-a-chip (LOC) community is to develop point-of-care diagnostic chips that do not use instruments. Such instruments include pumping or liquid handling devices for distribution of patient’s nucleic-acid test sample among an array of reactors and microvalves or mechanical parts to seal these reactors. In this paper, we report the development of a primer pair pre-loaded PCR array chip, in which the loading of the PCR mixture into an array of reactors and subsequent sealing of the reactors were realized by a novel capillary-based microfluidics with a manual two-step pipetting operations. The chip is capable of performing simultaneous (parallel) analyses of multiple gene targets and its performance was tested by amplifying twelve different gene targets against cDNA template from human hepatocellular carcinoma using SYBR Green I fluorescent dye. The versatility and reproducibility of the PCR-array chip are demonstrated by real-time PCR amplification of the BNI-1 fragment of SARS cDNA cloned in a plasmid vector. The reactor-to-reactor diffusion of the pre-loaded primer pairs in the chip is investigated to eliminate the possibility of primer cross-contamination. Key technical issues such as PCR mixture loss in gas-permeable PDMS chip layer and bubble generation due to different PDMS-glass bonding methods are investigated.
Starting Page	1007
Ending Page	1020
Page Count	14
File Format	PDF
ISSN	13872176
Journal	Biomedical Microdevices
Volume Number	11
Issue Number	5
e-ISSN	15728781
Language	English
Publisher	Springer US
Publisher Date	2009-05-07
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Capillary flow Capillary valve Microfluidic chip PCR array chip Real-time quantitative PCR Engineering Fluid Dynamics Nanotechnology Biophysics and Biological Physics Biomedical Engineering
Content Type	Text
Resource Type	Article
Subject	Nanoscience and Nanotechnology Molecular Biology Biomedical Engineering

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