Exploiting formalin‐preserved fish specimens for resources of DNA barcoding
Exploiting formalin‐preserved fish specimens for resources of DNA barcoding
复制标题
利用福尔马林保存的鱼类标本作为 DNA 条形码资源
DOI：
10.1111/j.1755-0998.2010.2838.x
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发表时间：
2010
影响因子：
7.7
通讯作者：
J. Zhang
中科院分区：
文献类型：
--
作者：
J. Zhang
随着DNA条形码项目的开展，需要大量的标本建立参考条形码库。来自博物馆的福尔马林固定样本为它提供了潜在的资源，然而从福尔马林固定样本中回收DNA和扩增目的基因具有挑战性。本研究采用热碱预处理结合十六烷基三甲基溴化铵（CTAB）法从福尔马林保存的样品中回收DNA，目的是寻求最佳条件以获得高数量和质量的DNA，并最大限度地减少PCR抑制。同时，开发了基于半巢式PCR的方法以提高扩增效率。这种先进的协议被证明是可靠和有效的。即使对于23岁的样本，也可以提取基因组DNA，并正确测序COI基因。
With the development of the DNA barcoding project, a large number of specimens are required to establish the library of reference barcode. Formalin‐fixed samples from museums provide a potential resource for it. However, recovery of DNA and amplification of the target gene from formalin‐fixed samples are challenging. In this study, a hot alkali pre‐treatment accompanied by the use of cetyltrimethylammonium bromide (CTAB) method was employed for DNA recovery from formalin‐preserved samples, with the purpose of pursuing the optimal condition for high quantity and quality of DNA and minimizing PCR inhibition. Meanwhile, a semi‐nested PCR‐based method was developed to enhance the efficacy of amplification. This advanced protocol was demonstrated to be reliable and effective. Even for 23‐year‐old samples, genomic DNA could be extracted, and COI gene was correctly sequenced.