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https://ptsldigital.ukm.my/jspui/handle/123456789/487029| Title: | Microfluidic device for DNA extraction and separation from saliva sample by superparamagnetic bead particles |
| Authors: | Samla Gauri Balakrishnan (P83770) |
| Supervisor: | Gan Kok Beng, Dr. |
| Keywords: | Microfluidic devices DNA Universiti Kebangsaan Malaysia -- Dissertations Dissertations, Academic -- Malaysia |
| Issue Date: | 18-Feb-2019 |
| Description: | The absence of impeccable information and primary empirical medication are the main factors which contribute to the increase of mortality rate in recent years. Bacteriology and pharmacogenomics studies are needed to minimize risk and complication associated with certain illness. Thus, personalized medicine based on the persons genetic profile such as DNA or genetic study is needed for tailored treatment plan without any side effect. In view of that, high throughput DNA extraction by superparamagnetic bead particles using microfluidic platform was studied using COMSOL Multiphysics® simulation software. Based on conceptual simulation study, microfluidic platform system for DNA extraction and separation from saliva sample is proposed as point of care device. The designed system was based on solid phase extraction using superparamagnetic bead particle in presence of electromagnetic field. The initial study covered effective DNA extraction using mobilizing superparamagnetic bead particles followed by DNA separation from superparamagnetic bead particles in electromagnetic field with appropriate elution buffer. MagJET magnetic bead particles were used to treat saliva sample for nucleic acid purification. The polycarbonate microfluidic channels consist of two parts which are micro-extraction and microseparation. Microfluidic micro mixer and superparamagnetic beads were used to extract DNA particles from saliva sample. The DNA particles which are attached with superparamagnetic beads are then transferred into continuous flow inline microchip for DNA purification. Based on results obtained from COMSOL Multiphysics® simulation, an average of 6 ng of DNA was extracted for every 1 microliter of sample input with 82.01% of purity. The system designed to illustrate this simulation consists of microcontroller, peristaltic pump, stepper motor, electromagnetic coil and others for fluid movement and DNA extraction. The proposed microfluidic platform together with electromagnetic field was able to extract and separate an average DNA concentration of 8.0 ng/µL with 1.8 (90%) purity from saliva sample using superparamagnetic bead particles. The extracted DNA was then quantified using NanoDrop™ spectrophotometer, amplified by PCR study and tested by gel electrophoresis analysis. The purified DNA appeared as positive PCR product at band 100 bp on the gel electrophoresis. Hence, a successful genomic DNA extraction form saliva sample was demonstrated. The microfluidic system with high stability of superparamagnetic beads provides an excellent chromatography separation medium for DNA sample preparation from saliva sample for further downstream analysis.,Ph.D. |
| Pages: | 137 |
| Call Number: | TJ853.4.M53S2395 2019 3 tesis |
| Publisher: | UKM, Bangi |
| Appears in Collections: | Faculty of Engineering and Built Environment / Fakulti Kejuruteraan dan Alam Bina |
Files in This Item:
| File | Description | Size | Format | |
|---|---|---|---|---|
| ukmvital_120993+SOURCE1+SOURCE1.1.PDF Restricted Access | 3.35 MB | Adobe PDF |  View/Open |
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