优化禽类羽毛中基因组 DNA 的提取:提高效率和成本效益的改良苯酚-氯仿法

IF 2.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Demir Ozdemir, Leyla Bener, Emine Toparslan Akcay
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引用次数: 0

摘要

在禽类物种中采集血液和组织样本提取 DNA 用于分子研究,会给研究对象带来不适和痛苦,再加上应用中的实际挑战和伦理方面的考虑。因此,羽毛已成为分子研究中更为普遍的来源,尤其是在家禽和鸟类学领域。然而,要从羽毛中有效提取 DNA,就必须分解羽毛结构中的硬角质化组织。本研究旨在设计一种高效、低成本、易适应的改良苯酚-氯仿(MPC)方法,用于从羽毛中提取基因组 DNA,以解决以往基于羽毛的 DNA 分离研究中发现的不足。我们采用 MPC 方法从六个不同禽类物种(鸡、珍珠鸡、金丝雀、鸽子、鸸鹋和鹅)的羽毛样本中提取基因组 DNA。在使用 MPC 方法的同时,还使用了两种不同的商用 DNA 试剂盒,对 DNA 分离效率进行了比较评估。结果显示,使用 MPC 方法从鸡毛中分离出的 DNA 浓度(纳克/毫升)明显高于使用商业试剂盒分离出的 DNA 浓度(p
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Optimizing Genomic DNA Extraction from Avian Feathers: A Modified Phenol-Chloroform Approach for Enhanced Efficiency and Cost-Effectiveness.

The procurement of blood and tissue samples for DNA extraction in avian species intended for molecular studies is associated with the induction of discomfort and pain in the subjects, compounded by practical challenges in application and ethical considerations. Consequently, feathers have emerged as a more prevalent source for molecular investigations, particularly in the fields of poultry and ornithology. However, the effective extraction of DNA from feathers necessitates the breakdown of the hard keratinized tissue within the feather structure. This study aimed to devise a highly efficient, cost-effective, and easily adaptable Modified Phenol-Chloroform (MPC) approach for genomic DNA extraction from feathers, addressing shortcomings identified in previous studies on feather-based DNA isolation. The MPC method was employed to extract genomic DNA from feather samples obtained from six distinct avian species (chicken, guinea fowl, canary, pigeon, emu, and goose). Comparative evaluation of DNA isolation efficiency was conducted by employing two different commercial DNA kits alongside the MPC method. The results showed significantly higher DNA concentrations (ng/ml) from chicken feathers using the MPC method compared to those obtained with commercial kits (p < 0.05), along with high DNA purity (1.83 ± 0.11). Subsequent PCR experiments, employing nuclear and mitochondrial DNA-specific primers, illustrated the effective amplification of short and long fragments from MPC-isolated DNA samples. In contrast to commercial kits, the findings underscore the successful application of the MPC method in isolating high-quality genomic DNA from feathers characterized by elevated keratin content.

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来源期刊
Biochemical Genetics
Biochemical Genetics 生物-生化与分子生物学
CiteScore
3.90
自引率
0.00%
发文量
133
审稿时长
4.8 months
期刊介绍: Biochemical Genetics welcomes original manuscripts that address and test clear scientific hypotheses, are directed to a broad scientific audience, and clearly contribute to the advancement of the field through the use of sound sampling or experimental design, reliable analytical methodologies and robust statistical analyses. Although studies focusing on particular regions and target organisms are welcome, it is not the journal’s goal to publish essentially descriptive studies that provide results with narrow applicability, or are based on very small samples or pseudoreplication. Rather, Biochemical Genetics welcomes review articles that go beyond summarizing previous publications and create added value through the systematic analysis and critique of the current state of knowledge or by conducting meta-analyses. Methodological articles are also within the scope of Biological Genetics, particularly when new laboratory techniques or computational approaches are fully described and thoroughly compared with the existing benchmark methods. Biochemical Genetics welcomes articles on the following topics: Genomics; Proteomics; Population genetics; Phylogenetics; Metagenomics; Microbial genetics; Genetics and evolution of wild and cultivated plants; Animal genetics and evolution; Human genetics and evolution; Genetic disorders; Genetic markers of diseases; Gene technology and therapy; Experimental and analytical methods; Statistical and computational methods.
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