Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Deutscher Platz 6, Leipzig, D-04103, Germany.
Wellcome Genome Campus, European Bioinformatics Institute (EMBL-EBI), Hinxton, Cambridgeshire, CB10 1SD, UK.
Biotechniques. 2021 Jul;71(1):382-386. doi: 10.2144/btn-2021-0025. Epub 2021 Jun 24.
Contamination with microbial and other exogenous DNA poses a significant challenge in the generation of genome-wide sequence data from ancient skeletal remains. Here we describe a method for separating ancient DNA into multiple fractions during DNA extraction by sequential temperature-controlled release of DNA into sodium phosphate buffer. An evaluation of the effectiveness of the method using a set of three ancient bones resulted in between 1.6- and 32-fold enrichment of endogenous DNA compared with regular DNA extraction. For two bones, the method outperformed previous methods of decontaminating ancient bones, including hypochlorite treatment, which resulted in near-complete destruction of DNA in the worst-preserved sample. This extraction method expands the spectrum of methods available for depleting contaminant DNA from ancient skeletal remains.
微生物和其他外源 DNA 的污染给从古代骨骼遗骸中生成全基因组序列数据带来了重大挑战。在这里，我们描述了一种在 DNA 提取过程中通过顺序温度控制将 DNA 释放到磷酸钠缓冲液中来将古代 DNA 分离成多个部分的方法。使用一组三个古代骨骼对该方法的有效性进行评估，与常规 DNA 提取相比，内源性 DNA 的富集倍数在 1.6 到 32 倍之间。对于两个骨骼，该方法优于以前的古代骨骼去污方法，包括次氯酸钠处理，该方法导致保存最差的样本中 DNA 几乎完全破坏。这种提取方法扩大了从古代骨骼遗骸中去除污染物 DNA 的方法的范围。
