School of Biological Sciences, University of Sydney, Sydney, NSW 2006, Australia.
J Mol Evol. 2011 Oct;73(3-4):244-55. doi: 10.1007/s00239-011-9474-z. Epub 2011 Nov 20.
The growth of ancient DNA research has offered exceptional opportunities and raised great expectations, but has also presented some considerable challenges. One of the ongoing issues is the impact of post-mortem damage in DNA molecules. Nucleotide alterations and DNA strand breakages lead to a significant decrease in the quantity of DNA molecules of useful length in a sample and to errors in the final DNA sequences obtained. We present a model of age-dependent DNA damage and quantify the influence of that damage on subsequent steps in the sequencing process, including the polymerase chain reaction and cloning. Calculations using our model show that deposition conditions, rather than the age of a sample, have the greatest influence on the level of DNA damage. In turn, this affects the probability of interpreting an erroneous (possessing damage-derived mutations) sequence as being authentic. We also evaluated the effect of post-mortem damage on real data sets using a Bayesian phylogenetic approach. According to our study, damage-derived sequence alterations appear to have little impact on the final DNA sequences. This indicates the effectiveness of current methods for sequence authentication and validation.
古 DNA 研究的发展提供了特殊的机会和很高的期望,但也带来了一些相当大的挑战。其中一个持续存在的问题是 DNA 分子死后损伤的影响。核苷酸的改变和 DNA 链的断裂导致样本中有用长度的 DNA 分子数量显著减少,并导致最终获得的 DNA 序列出现错误。我们提出了一个年龄相关的 DNA 损伤模型,并量化了这种损伤对测序过程中后续步骤的影响,包括聚合酶链反应和克隆。使用我们的模型进行的计算表明,沉积条件而不是样本的年龄对 DNA 损伤水平有最大的影响。反过来,这又会影响将错误的(具有损伤衍生突变的)序列解释为真实序列的概率。我们还使用贝叶斯系统发育方法评估了死后损伤对真实数据集的影响。根据我们的研究,损伤衍生的序列改变似乎对最终的 DNA 序列影响不大。这表明当前用于序列验证和确认的方法是有效的。