Comparative Evaluation of Different Established Genomic Extraction Methods for Scleractinian Coral

Q4 Multidisciplinary
Muhammad Irsyad Abiyusfi Ghafari, R. Agus, M. Litaay
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引用次数: 0

Abstract

Studies on coral genetics have been widely used recently for varied research purposes, including to solve the taxonomical challenge or to understand the physiological-ecological aspects of corals towards climate change. While the information provided by coral genome is crucial for those works, isolation of coral genome is still a difficult preliminary step due to coral's nature. This study aimed to evaluate and to suggest the best application of among four established methods for DNA extraction from scleractinian coral, i.e., Guanidine hydrochloride (GuHCl), Chelex chelating resin, Guanidinium thiocyanate-phenol-chloroform (AGPC), and spin-column methods. Coral samples were grounded and submitted to each extraction method's protocol. Final extracts were checked for their quality and quantity. The final extracts feasibility were tested through PCR (Polymerase Chain Reaction), followed by electrophoresis for amplicon quality screening. The scoring was done for assessment aspects: time efficiency, successful extraction process, extracted DNA quantity and quality, final extract properness, and cost effort per sample. The highest DNA concentration was obtained by spin-column methods, reaching up to 724.31 ng/µl, while other methods can only provide between 264.94±36.453 to 378.19±0 ng/µl of DNA. Among all method tested, the spin-column was able to provide reproducible and feasible quality of final extracts for the amplification. Other methods were failed to provide such consistent and proper results. The results suggesthat the spin-column is the most appropriate method for DNA isolation from scleractinian specimen.
硬核珊瑚不同基因组提取方法的比较评价
珊瑚遗传学研究最近被广泛用于各种研究目的,包括解决分类学挑战或了解珊瑚对气候变化的生理生态方面。虽然珊瑚基因组提供的信息对这些工作至关重要,但由于珊瑚的性质,分离珊瑚基因组仍然是一个困难的初步步骤。本研究旨在评估并建议四种已建立的巩膜珊瑚DNA提取方法中的最佳应用,即盐酸胍(GuHCl)、螯合树脂、硫氰酸胍苯酚-氯仿(AGPC)和旋转柱法。珊瑚样品被固定并提交给每种提取方法的方案。检查最终提取物的质量和数量。通过PCR(聚合酶链式反应),然后通过电泳进行扩增子质量筛选,测试最终提取物的可行性。对评估方面进行评分:时间效率、成功提取过程、提取的DNA数量和质量、最终提取的适宜性以及每个样本的成本努力。通过旋转柱法获得的DNA浓度最高,达到724.31纳克/µl,而其他方法只能提供264.94±36.453至378.19±0纳克/µl的DNA。在所有测试的方法中,旋转柱能够为扩增提供可重复和可行的最终提取物质量。其他方法未能提供如此一致和适当的结果。结果表明,旋转柱是从巩膜标本中分离DNA最合适的方法。
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来源期刊
ASM Science Journal
ASM Science Journal Multidisciplinary-Multidisciplinary
CiteScore
0.60
自引率
0.00%
发文量
23
期刊介绍: The ASM Science Journal publishes advancements in the broad fields of medical, engineering, earth, mathematical, physical, chemical and agricultural sciences as well as ICT. Scientific articles published will be on the basis of originality, importance and significant contribution to science, scientific research and the public. Scientific articles published will be on the basis of originality, importance and significant contribution to science, scientific research and the public. Scientists who subscribe to the fields listed above will be the source of papers to the journal. All articles will be reviewed by at least two experts in that particular field.
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