Jasmine Eva, Ajan (2018) Assessing Escherichia coli Genomic DNA Integrity during Extraction Using Selected Protocols. [Final Year Project Report] (Unpublished)
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Abstract
Escherichia coli (E. coli) is a common inhabitant of the gastrointestinal tracts of humans and warm-blooded animals. E. coli is frequently used as a model organism in molecular biology and microbiology studies due to its peculiar characteristics. Thus, its genomic deoxyribonucleic acid (DNA) is frequently extracted by using different types of DNA extraction protocols in order to study its genomic sequence. Thus, a rapid and costefficient genomic DNA extraction protocol is essential for the purpose of molecular biology studies, and extraction of high quality and quantity genomic DNA. In this study, four common DNA extraction methods were used for E. coli DNA extraction, namely, boil cell method, phenol-chloroform isoamyl alcohol (PCI) method, and commercial DNA extraction kit with and without bead beatings methods. The DNA extracted from each protocol was analysed by using a SynergyTM HI system and M13-Polymerase Chain Reaction (PCR) and Random Amplified Polymorphic DNA (RAPD)-PCR fingerprinting. DNA yield and purity were quantified using SynergyTM HI system absorbance ratios. The extracted DNA was also analyzed using agarose gel electrophoresis. Results show differences in DNA yield, purity and PCR success between extraction methods. All four methods tested provided appropriate DNA for PCR amplification, but with different yields. In this study, the commercial DNA extraction kit without bead-beating method was found to be the most efficient DNA extraction method, capable to provide high DNA yields with better quality, affordable cost and less time.
| Item Type: | Final Year Project Report |
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| Additional Information: | Project Report (B.Sc.) -- Universiti Malaysia Sarawak, 2018. |
| Uncontrolled Keywords: | Escherichia coli, genomic DNA, DNA extraction method, M13-PCR, RAPD-PCR, DNA yield, DNA purity |
| Subjects: | Q Science > Q Science (General) |
| Divisions: | Academic Faculties, Institutes and Centres > Faculty of Resource Science and Technology Faculties, Institutes, Centres > Faculty of Resource Science and Technology |
| Depositing User: | Unai |
| Date Deposited: | 30 Sep 2021 04:21 |
| Last Modified: | 23 Jun 2023 09:33 |
| URI: | http://ir.unimas.my/id/eprint/36238 |
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