Alberts C C, Ribeiro-Paes J T, Aranda-Selverio G, Cursino-Santos J R, Moreno-Cotulio V R, Oliveira A L D, Porchia B F M M, Santos W F, Souza E B
Departamento de Ciências Biológicas, Faculdade de Ciências e Letras de Assis, Universidade Estadual Paulista, Assis, SP, Brasil.
Genet Mol Res. 2010 Dec 21;9(4):2429-35. doi: 10.4238/vol9-4gmr1027.
Wild felids and canids are usually the main predators in the food chains where they dwell and are almost invisible to behavior and ecology researchers. Due to their grooming behavior, they tend to swallow shed hair, which shows up in the feces. DNA found in hair shafts can be used in molecular studies that can unravel, for instance, genetic variability, reproductive mode and family structure, and in some species, it is even possible to estimate migration and dispersion rates in given populations. First, however, DNA must be extracted from hair. We extracted successfully and dependably hair shaft DNA from eight wild Brazilian felids, ocelot, margay, oncilla, Geoffroy's cat, pampas cat, jaguarundi, puma, and jaguar, as well as the domestic cat and from three wild Brazilian canids, maned wolf, crab-eating fox, and hoary fox, as well as the domestic dog. Hair samples came mostly from feces collected at the São Paulo Zoo and were also gathered from non-sedated pet or from recently dead wild animals and were also collected from museum specimens. Fractions of hair samples were stained before DNA extraction, while most samples were not. Our extraction protocol is based on a feather DNA extraction technique, based in the phenol:chloroform:isoamyl alcohol general method, with proteinase K as digestive enzyme.
野生猫科动物和犬科动物通常是它们所处食物链中的主要捕食者，行为和生态研究人员几乎难以观察到它们。由于它们的梳理行为，它们往往会吞下脱落的毛发，这些毛发会出现在粪便中。毛干中发现的DNA可用于分子研究，例如揭示遗传变异性、繁殖方式和家族结构，对于某些物种，甚至可以估计特定种群中的迁移和扩散率。然而，首先必须从毛发中提取DNA。我们成功且可靠地从八种巴西野生猫科动物（豹猫、虎猫、小斑虎猫、乔氏猫、潘帕斯猫、美洲獭猫、美洲狮和美洲豹）、家猫以及三种巴西野生犬科动物（鬃狼、食蟹狐和苍狐）、家犬中提取了毛干DNA。毛发样本大多来自圣保罗动物园收集的粪便，也从未镇静的宠物、刚死亡的野生动物以及博物馆标本中采集。部分毛发样本在DNA提取前进行了染色，而大多数样本未染色。我们的提取方案基于一种羽毛DNA提取技术，该技术基于酚：氯仿：异戊醇通用方法，以蛋白酶K作为消化酶。
