用于大鼠 PCR 基因分型的直肠拭子 DNA 采集方案。

IF 2.2 4区工程技术 Q3 BIOCHEMICAL RESEARCH METHODS

BioTechniques Pub Date : 2024-04-23 DOI:10.2144/btn-2024-0023

Audrey E. Kaye, Jacob W. Proctor-Bonbright, Jai Y. Yu

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引用次数: 0

摘要

DNA 采集对实验室动物基因分型至关重要。常见的采集方法需要切除组织，会造成不适和损伤。直肠拭子采集法被认为是一种有效的微创替代方法，但目前还没有证据支持该技术的方案。本报告评估了采集参数对 PCR 结果质量的影响，并提出了在 3-5 小时内对一窝大鼠进行基因分型的方案。2-8 次刮取的样本可产生足够的 DNA，利用 PCR 可扩增出长达∼1800 bp 的目标。直肠拭子产生的 PCR 结果与耳夹样本的效用相似，而且结果不受残留粪便或细胞碎片的影响。该方案可使用商用 PCR 试剂进行快速、微创和可重复的基因分型。

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Rectal swab DNA collection protocol for PCR genotyping in rats.

DNA collection is essential for genotyping laboratory animals. Common collection methods require tissue amputation, causing discomfort and injury. Rectal swabbing has been proposed as an effective, minimally invasive alternative, but an evidence-backed protocol for the technique remains unavailable. This report evaluates the effects of collection parameters on the quality of PCR results and presents a protocol for genotyping a litter of rats within 3-5 h. Samples with 2-8 scrapes produced enough DNA to amplify targets up to ∼1800 bp long using PCR. Rectal swabbing produced PCR results with similar utility as ear clip samples, and results were unaffected by residual fecal matter or cell debris. The protocol enables fast, minimally invasive and repeatable genotyping using commercial PCR reagents.

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来源期刊

BioTechniques 工程技术-生化研究方法

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

3.3 months

期刊介绍： BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.