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Alternatives to Blood as a Source of DNA for Large-scale Scanning Studies of Canine Genome Linkages

机译:犬基因组链接的大规模扫描研究中血液作为DNA来源的替代方法

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Participation and compliance are critical to the success of any large-scale study of canine disease using DNA markers. Most canine genetic studies rely upon DNA extracted from peripheral blood samples. We assessed the utility of buccal swab epithelial cells and toe nails as a source of DNA for use in genomic screening studies. Using eight multiplexed canine microsatellite markers, amplified DNA obtained from peripheral blood, and from freshly extracted buccal epithelial cells, and buccal swab DNA extracted and stored at –20°C for 27 months or extracted from toe nails were compared for three dogs. The accuracy of the genotyping at each locus was identical for each preparation. Buccal swab DNA samples were readily and uniformly amplified and could be stored for years without loss of integrity. Each buccal swab provided sufficient DNA for more than 200 individual PCR reactions. Toe nails provided ample DNA for thousands of PCR reactions and had the added advantage of ease of storage of the original tissues. These studies demonstrate the potential utility of DNA derived from buccal swabs or nails in large-scale genomic scanning and marker linkage studies.
机译:参与和依从性对于使用DNA标记物进行的犬病大规模研究的成功至关重要。大多数犬类遗传学研究都依赖于从外周血样本中提取的DNA。我们评估了口腔拭子上皮细胞和脚趾指甲作为用于基因组筛选研究的DNA来源的实用性。使用八种多重犬微卫星标记,比较了三只狗从外周血和新鲜提取的颊上皮细胞中获得的扩增DNA,以及在–20°C下提取并存储在27个月或从脚趾指甲中提取的颊拭子DNA。对于每种制剂,每个基因座处基因分型的准确性均相同。颊拭子DNA样本易于均匀地扩增,可以保存多年而不失完整性。每个颊拭子为200多个单独的PCR反应提供了足够的DNA。脚趾指甲可为数千个PCR反应提供充足的DNA,并具有易于保存原始组织的额外优势。这些研究证明了从口腔拭子或指甲中提取的DNA在大规模基因组扫描和标记连锁研究中的潜在用途。

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