甲醛溶液保存的中华哲水蚤基因组DNA提取和PCR扩增  

Extraction of Genomic DNA and Amplification of mtDNA COI from Calanus sinicus Stored in Formaldehyde Solution

作　　者：周美玉[1] 林元烧[1] 方旅平[1] 郑连明[1] 刘迟迟[1] 曹文清[1] 

机构地区：[1]厦门大学海洋与环境学院,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2005年第3期403-406,共4页Journal of Xiamen University：Natural Science

基　　金：国家重点基础研究发展规划 ( 973 )项目资助(G1999043708)

摘　　要：提取了保存于5%甲醛溶液中的单只中华哲水蚤基因组DNA.经凝胶电泳后虽无清晰条带,但PCR扩增后可得线粒体DNA细胞色素氧化酶I亚基基因(mtCOI)片段.测序分析,该片段长度为710bp,与GenBank中的序列(索引号AF332769)比对后确定为中华哲水蚤mtCOI序列的一部分.利用本实验方法所提取的基因组DNA可适用于系统发生、种群遗传结构等领域的分子水平研究,为保存在甲醛溶液中的小型海洋浮游动物的基因信息利用提供实用技术.The DNA extraction,in General,from samples stored in formaldehyde solution is inefficient,especially extracted from small individual.It is necessary extracting of DNA genome from formaldehyde samples which conserved in early days or fixed in formaldehyde solution which usually adopted in marine investigation. On the base of methods of DNA extraction used in other animals or tissues,genomic DNA is extracted from Calanus sinicus stored in 5% formaldehyde solution in this experiment.More attention was paid to the process of eliminating the formaldehyde solution at the very beginning and at the digestion period,we add DTT into the digestion reaction systemwhich are the most important steps in the whole experiment.The concentration of genomic DNA in the study is low relatively,but after efficient PCR amplification,mtDNA COI gene fragment (710 bp) was obtained.Then comparing with the sequence of Calanus sinicus recorded in Genbank (accession no.AF332769),it is sure of that what we get covers the recorded of GenBank,is Calanus sinicus mtDNA COI sequence.The study will serve other small pelagic crustacean for the extracting DNA genome.It can also apply to the research of phylogeny,the population genetics and so on at molecular level,providing the use of gene information of small marine zooplankton samples stored in formaldehyde solution with practical technique.

关 键 词：中华哲水蚤 PCR扩增 基因组DNA提取 甲醛溶液 保存 细胞色素氧化酶 GENBANK 线粒体DNA 海洋浮游动物 种群遗传结构 测序分析 凝胶电泳 片段长度 系统发生 实验方法 实用技术 信息利用 分子水平 序列 

分 类 号：Q959.223.3[生物学—动物学] Q78