Automated,high-throughput DNA extraction protocol for disposable label free,microfluidics integrating DNA biosensor for oil palm pathogen,Ganoderma boninense |
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| Institution: | 1. Ganoderma Research Group, Institute of Nano Electronic Engineering (INEE), Universiti Malaysia Perlis, 01000, Kangar, Malaysia;2. School of Bioprocess Engineering, Universiti Malaysia Perlis, 02600, Arau, Perlis, Malaysia;1. Division of Engineering in Medicine, Department of Medicine, Brigham and Women''s Hospital – Harvard Medical School, Boston, MA 02115, USA;2. Center for Nanotechnology, King AbdulAziz University, Jeddah 21589, Saudi Arabia;3. Division of Infectious Disease, Department of Medicine, Brigham and Women''s Hospital – Harvard Medical School, Boston, MA 02115, USA;4. Department of Medicine, Harvard Medical School, 25 Shattuck Street, Boston, MA 02115, USA;1. NTT Hi-Tech Institute, Nguyen Tat Thanh University, Ho Chi Minh City, Viet nam;2. Department of Chemistry, Pukyong National University, Busan, 608-737, Republic of Korea;3. Department of Biochemistry, Faculty of Biology and Biotechnology, University of Science, Vietnam National University, Ho Chi Minh City, Viet nam;4. Faculty of Natural Sciences, Thu Dau Mot University, Thu Dau Mot City, Binh Duong Province, Viet nam;1. Department of Biocatalysis, Institute of Catalysis and Petrochemistry, Consejo Superior de Investigaciones Científicas (CSIC), 28049 Madrid, Spain;2. Department of Chemistry, Universidad de Guadalajara, 44430 Jalisco, Mexico;3. Department of Chemistry, Kharazmi University, 1417466191 Tehran, Iran;4. Pharmacy and Biotechnology Department, School of Biomedical Sciences, Universidad Europea de Madrid, Villaviciosa de Odón, 28670 Madrid, Spain;5. Laboratory of Microbiology and Food Biocatalysis, Institute of Food Science Research (CIAL, CSIC-UAM). Nicolás Cabrera, 9. UAM Campus, Cantoblanco, 28049, Madrid, Spain;1. School of Life Sciences, Ludong University, Yantai 264025, China;2. School of Agriculture, Ludong University, Yantai 264025, China |
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| Abstract: | Basal Stem Rot and Upper Stem Rot diseases caused by pathogenic fungus Ganoderma boninense continue to be a major plight in the palm oil industry. Despite continuous research in combating the problem, resolution remains stagnant. Here, developed an automated, high-throughput DNA extraction protocol on microfluidics device for a quick, disposable, label-free detection, within 2 h of assessment. Microfluidics was designed using AutoCAD software, fabricated on microscopic glass substrate using negative photoresist (SU-8 2015) and molded with a biopolymer silicone, Polydimethylsiloxane. G. boninense and unknown pathogenic fungus isolated from rotten mushroom were grown and fractions of extracted DNA were pooled and analyzed for comparison along with synthetic ssDNA of G. boninense. Results from LPM and HPM show successful fabrication with ≤0.1 mm variance between the dimensions in the design before and after lithography process. The PDMS microfluidics show no leakage when run with DNA samples. Analyses from I-V measurement, UV–vis, FTIR, and PCR show comparable results between extracted and synthetic ssDNA of G. boninense and a contrast with the unknown pathogenic fungus, indicating a successful DNA extraction protocol via microfluidics for label-free identification of G. boninense. Optimization of DNA extraction can be further devised for applicability on lab-on-a-chip devices. |
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| Keywords: | DNA extraction Polydimethylsiloxane Microfluidics Biosensor |
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