Non-random DNA damage resulting from heat treatment: implications for sequence analysis of ancient DNA
Non-random DNA damage resulting from heat treatment: implications for sequence analysis of ancient DNA
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DOI：
10.1016/s0305-4403(03)00099-2
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发表时间：
2004-01-01
影响因子：
2.8
通讯作者：
Brown, TA
中科院分区：
文献类型：
--
作者：
Banerjee, M;Brown, TA
为了模拟古代DNA保存过程中发生的损伤，我们将小麦DNA在95 ℃下加热2-21天。通过针对线粒体atpA基因的181-bp片段的PCR扩增样品。对PCR产物进行克隆，获得多个序列。我们预期表明损伤的核苷酸变化将随机分布，但情况并非如此。在124个克隆的序列中，107个显示出相同的腺嘌呤到鸟嘌呤的变化，这种变化在来自所有8个加热样品的克隆中均可见。来自6个样品的15个克隆序列在位置92处显示出相同的明显缺失。如果非随机序列变化是DNA降解的一个普遍特征，那么必须修改当前用于分析古DNA序列的策略，因为这些策略可能会导致核苷酸变异，这些变异是由DNA成岩作用引起的人工制品，被误认为是古DNA的真正序列特征。(C)2003爱思唯尔有限公司。保留所有权利。
To mimic damage occurring during preservation of ancient DNA, we heated wheat DNA at 95 degreesC for 2-21 days. Samples were amplified by PCRs directed at a 181-bp segment of the mitochondrial atpA gene. PCR products were cloned and multiple sequences obtained. We anticipated that nucleotide changes indicating damage would be randomly distributed, but this was not the case. Of the 124 cloned sequences, 107 displayed the same adenine to guanine change, this change being seen in clones from all of the eight heated samples. Fifteen of the cloned sequences, from six samples, showed the same apparent deletion at position 92. If non-random sequence changes are a widespread feature of DNA degradation then current strategies for analysis of ancient DNA sequences must be revised, as these could lead to nucleotide variations that are artefacts resulting from DNA diagenesis being mistaken for genuine sequence features of the ancient DNA. (C) 2003 Elsevier Ltd. All rights reserved.