Rohland Nadin, Siedel Heike, Hofreiter Michael
Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany.
Biotechniques. 2004 May;36(5):814-6, 818-21. doi: 10.2144/04365ST05.
Museum specimens have provided the material for a large proportion of ancient DNA studies conducted during the last 20 years. However, a major drawback of the genetic analyses is that the specimens investigated are usually damaged, as parts of skin, bone, or a tooth have to be removed for DNA extraction. To get around these limitations, we have developed a nondestructive extraction method for bone, tooth, and skin samples. We found that it is possible to amplify mitochondrial DNA (mtDNA) sequences up to at least 414 bp long from samples up to 164 years old. Using this method, almost 90% (35 of 40) of the investigated samples yielded amplifiable mtDNA. Moreover, we found that repeated extractions of the same samples allowed amplifications of the expected length for all samples at least three times and for some samples up to at least five times. Thus this method opens up the possibility to repeatedly use museum collections for mtDNA analyses without damaging the specimens and thus without reducing the value of irreplaceable collections for morphological analyses.
在过去20年里,博物馆标本为大部分古代DNA研究提供了材料。然而,基因分析的一个主要缺点是,所研究的标本通常已受损,因为为了提取DNA,必须去除部分皮肤、骨骼或牙齿。为了克服这些限制,我们开发了一种针对骨骼、牙齿和皮肤样本的非破坏性提取方法。我们发现,从年龄达164岁的样本中,有可能扩增出长度至少达414 bp的线粒体DNA(mtDNA)序列。使用这种方法,近90%(40个样本中的35个)被研究样本产生了可扩增的mtDNA。此外,我们发现对同一样本进行重复提取能使所有样本至少三次、有些样本至少五次扩增出预期长度。因此,这种方法开启了在不损坏标本的情况下反复使用博物馆藏品进行mtDNA分析的可能性,从而不会降低不可替代藏品用于形态学分析的价值。