Rectal Swab DNA Collection Protocol for PCR Genotyping in Rats.
Rectal Swab DNA Collection Protocol for PCR Genotyping in Rats.
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用于大鼠 PCR 基因分型的直肠拭子 DNA 采集方案。
DOI：
10.1101/2024.03.02.583131
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发表时间：
2024
期刊：
影响因子：
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通讯作者：
Yu,JaiY
中科院分区：
文献类型：
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作者：
Kaye,AudreyE;Proctor-Bonbright,JacobW;Yu,JaiY
DNA收集对于对实验动物进行基因分型是必不可少的。常见的收集方法需要截断组织，导致不适和受伤。直肠拭子已经被认为是一种有效的、微创的替代方法，但该技术的证据支持的方案仍然不可用。这份报告评估了收集参数对聚合酶链式反应结果质量的影响，并提出了一种在3-5小时内对一窝大鼠进行基因分型的方案。用2-8个刮片的样本产生的DNA足以用聚合酶链式反应扩增长达1800bp的∼。直肠拭子产生的聚合酶链式反应结果与耳夹样本类似，结果不受残留粪便或细胞碎片的影响。该协议允许使用商业PCR试剂进行快速、微创检测和可重复的基因分型。
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.