%0 Journal Article
%T Discussing on the feasibility of large fragments DNA extraction from formalin-fixed fish specimens<br>福尔马林保存鱼类标本中大片段DNA的提取
%A XIA Ying-Zhe
%A ZHANG Chun-Guang
%A CHEN Yi-Yu
%A SHENG Yan
%A SUN Yue-Hua
%A <br>夏颖哲
%A 张春光
%A 陈宜瑜
%A 盛岩
%A 孙悦华
%J 水生生物学报
%D 2007
%I 
%X Formalin-fixed specimens were of profound importance to the study of evolution,systematic,phylogeny and genetic structure of populations.However,the PCR analysis of DNA isolated from formalin-fixed tissue could be difficult because of the inherently poor quality of template DNA available for amplification.As a consequence of fixation process,DNA was complex with proteins and is often nicked,giving relatively short fragments.Such samples were often of low DNA concentration and poor quality.Formalin-fixation had a profound effect on the molecular arrangement of nucleic acids in the tissue and resulted almost invariably in DNA degradation of various degrees.Extraction,purification and amplification of DNA from formalin-fixed tissues were influenced by a multitude of factors.In this report,we introduced a method of DNA extraction and PCR amplification of relatively large target fragments from formalin-fixed,fluid-preserved tissues.Main modifications include: long-time preservation in buffer,short-time heating,and drying in a vacuum centrifuge during pre-disposal;more proteinase K and sodium SDS during extraction;performing PCR immediately after digestion,prolonging annealing time,and increasing the number of cycles.The feasibility of this method was tested by PCR amplification.The nucleotide DNA of cloned products from formalin-fixed specimens were sequenced and the results were compared with the sequences obtained from fresh and ethanol-fixed tissue and published data.Comparison of the DNA sequence data from the formalin-fixed tissues with that from the frozen and ethanol-fixed tissues demonstrated this method is reliable.We also found that the pre-fixation time could be an important factor determining the quality of DNA.
%K DNA<br>福尔马林
%K 固定标本
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=256651267D5D2E4520A7707D05E98747&aid=1BD1426169E2E9BD&yid=A732AF04DDA03BB3&vid=4AD960B5AD2D111A&iid=E158A972A605785F&sid=CA4FD0336C81A37A&eid=ABF2590617D31FFD&journal_id=1000-3207&journal_name=水生生物学报&referenced_num=0&reference_num=29