Qualitative comparison of DNA isolation methods from insects specimen

Rodrigo Ordoñez

Bachelor's thesis

Abstract:

This study compares the performance of different molecular methods, including DNA extraction kits and direct PCR in biodiversity research using the specimen model Thanatophilus sinuatus. By determining the efficacy and accuracy of these molecular techniques, we aim to identify the most effective method for enhancing the proper identification of targeted species. The results of this study will provide valuable insights into the strengths and limitations of each method and their future implications in the biodiversity research study field. DNA was extracted from specimens of Thanatophilus sinuatus using a commercial DNA extraction kit (Genomic DNA Mini Kit) and also performed direct PCR amplification on the same specimens without prior extraction. The efficacy and accuracy of these two molecular methods was compared in terms of DNA yield, quality, and suitability for downstream analyses, including PCR amplification and sequencing. The DNA was later amplified and sequenced the cytochrome c oxidase subunit I (COI) gene from each sample, and then compared the quality and reliability of the resulting sequences. Also the cost-effectiveness and time-efficiency of the two methods was compared. The results suggest that direct PCR amplification is a cost-effective and time-efficient alternative to commercial DNA extraction kits (Genomic DNA Mini Kit) for biodiversity research. Direct PCR produced PCR and sequencing results that were comparable to those obtained using the DNA extraction kit. Furthermore, direct PCR amplification was significantly cheaper and faster than the DNA extraction kit, making it a more practical and efficient molecular method for biodiversity research. The results of this study shows the reliability of Direct PCR method as a cost-effective and time-efficient alternative to the commercial DNA extraction kit used, for biodiversity research, giving out a valuable contribution to the field of biodiversity research studies.

Keywords

DNA, Isolation, Entomology

Language used: English

Date on which the thesis was submitted / produced: 4. 4. 2023

Thesis defence

• Date of defence: 24. 5. 2023
• Supervisor: Ing. Pavel Jakubec, Ph.D.
• Reader: Jarin Qubaiová, Ph.D. MSc

Czech University of Life Sciences Prague

Faculty of Environmental Sciences

Bachelor programme:
Environmental Engineering