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首页> 外文期刊>Wildlife Society Bulletin >Use of Buccal Swabs for Sampling DNA from Nestling and Adult Birds
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Use of Buccal Swabs for Sampling DNA from Nestling and Adult Birds

机译:颊拭子用于从雏鸟和成年鸟中采集DNA的采样

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We evaluated the feasibility and efficiency of using swabs to collect buccal epithelial cells from small (2- to 13-g) birds as a source of DNA for genetic studies. We used commercially available buccal swab kits to collect samples from 42 adult and 39 nestling (4- to 8-day-old) black-capped chickadees (Poecile atricapillus) and from 6 4-day-old nestling boreal chickadees (P. hudsonica). We compared DNA from buccal epithelial samples to that from blood samples from the same individuals. We extracted sufficient quantities of DNA for analysis from all buccal samples, and samples remained viable even after being stored in original plastic sampling tubes at room temperature for up to 18 months. Yields were equivalent whether extracted using the proprietary quick-extraction solution provided with buccal swab kits or using a salt-extraction process with inexpensive reagents. Yields of DNA from buccal samples were consistently lower than those from blood samples, but quantities were sufficient for all analyses. Assignment of sex, based on DNA extracted from paired buccal and blood samples, was identical for all 87 birds. We found no difference in the genotypes obtained from buccal and blood samples for 12 individuals tested using 5 microsatellite loci and found perfect concordance in sequencing of an 823-base-pair segment within the control region of mitochondrial DNA for 7 individuals tested. Use of buccal swabs is highly recommended as a rapid, noninvasive technique for sampling avian genomic DNA, especially for extremely young altricial nestlings or small-bodied adults, or for any birds for which blood sampling may be impossible or stressful.
机译:我们评估了使用拭子收集小鸟(2至13克)作为基因研究DNA来源的颊上皮细胞的可行性和效率。我们使用市售的口腔拭子试剂盒收集了42例成年和39只雏鸟(4至8天大)黑冠山雀(Poecile atricapillus)和6只4天龄的雏鸟北半山雀(P. hudsonica)的样本。我们比较了颊上皮样品的DNA和同一个人血液样品的DNA。我们从所有颊样品中提取了足够量的DNA进行分析,即使将样品在原始塑料采样管中于室温下保存长达18个月之后,它们仍然具有活力。无论是使用颊拭子套件随附的专有快速萃取溶液进行萃取,还是使用廉价试剂进行盐萃取过程,产量均相等。颊样品中DNA的产量始终低于血液样品中的DNA,但数量足以进行所有分析。根据从成对的颊和血液样本中提取的DNA进行的性别分配,对于所有87只鸡来说都是相同的。我们发现使用5个微卫星基因座测试的12位个体从颊和血液样本获得的基因型没有差异,并且在7位测试的线粒体DNA控制区域内的823个碱基对片段的测序中发现了完全一致。强烈建议使用口腔拭子作为一种快速,无创的​​技术来采集禽类基因组DNA,特别是对于幼小的幼鸟雏鸟或成年成年幼鸟,或者对无法进行血液采样或压力很大的鸟类而言。

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