福尔马林固定标本DNA提取及富集实验方法优化

DNA Extraction and Enrichment From Formalin-fixed Specimens Experimental Method Optimization

早期鱼类标本的保藏,为了防止标本变形利于后续研究,对稀少珍贵的鱼类常采用福尔马林浸泡的方法。但是经过长期的保存,由于基因与基因、蛋白质与蛋白质、基因与蛋白质之间发生交联作用,导致提取长片段基因组DNA的难度增大,通过PCR方法获取目标物种基因序列尤其是长片段存在诸多困难。本文通过文献查阅及实验方法总结,并在实验室的条件下对提取方法进行优化和后续实验验证,获得一种比较方便有效的实验方法。根据优化后的实验方法在实验室条件下获得了相应的小片段DNA序列,对所获得的片段进行BLAST分析和系统发育树,证实所获得的片段为目标物种序列,证明该实验方法具备可行性,可用于福尔马林长期固定标本DNA提取,以获取目的片段。

In order to prevent specimen deformation and facilitate subsequent research, formalin soaking method is often used for rare and precious fish, in the preservation of early fish specimens. However, after longterm preservation, due to the crosslinking between genes and genes, proteins and proteins, genes and proteins, it is more difficult to extract long-fragment genomic DNA, and there are many difficulties in obtaining the gene sequences of the target species, especially long fragments, by PCR methods. In this paper, a more convenient and effective experimental method is obtained. through literature review and summary of experimental methods, and the optimization of extraction methods and subsequent experimental verification under laboratory conditions. According to the optimized experimental method, the corresponding small fragments of DNA sequences were obtained under laboratory conditions, and the obtained fragments were analyzed and phylogenetic trees were performed to confirm that the obtained fragments were target species sequences, which proved that the experimental method was feasible and could be used for long-term fixed specimen DNA extraction of formalin to obtain the fragment of interest.