鸟类陈旧标本DNA提取方法的研究  

Methods of DNA Extraction from Old Specimen of Wetland Bird

作　　者：陈小麟[1] 麻常昕[1] 李琪[1] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2008年第A02期1-5,共5页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金项目(30570254);福建省科技厅科技计划重点项目(2008S0007)

摘　　要：应用3种DNA提取方法(蛋白酶K法,Chelex 100法,硅颗粒法),从馆藏湿地鸟类陈旧标本的羽毛、皮肤、骨骼组织中提取基因组DNA,经PCR扩增后对线粒体12S rDNA部分片段进行测序和Genbank查对,以探讨鸟类陈旧标本DNA的最佳提取方法和提取组织.结果表明,利用鸟类陈旧标本能够提取到基因组DNA.蛋白酶K法和硅颗粒法能够从白鹭(Egretta garzetta)标本的骨骼组织中提取到DNA并获得长度425 bp的线粒体12S rDNA序列(Genbank接收号AY766387),蛋白酶K法还能够从白鹳(C iconia ciconia)标本的羽毛中获得长度263 bp的线粒体12S rDNA序列(Gen-bank接收号AY766388),Chelex 100法不能从鸟类标本中提取到DNA.蛋白酶K法的DNA提取量较大,但是步骤繁琐而导致DNA被污染的几率增大,因此,实验过程中要注意防止污染,且提取骨组织DNA要经过纯化.硅颗粒法的步骤简单,污染几率降低,提取骨骼组织DNA不需经过纯化,但是其DNA提取量较少,不能用于羽毛DNA的提取.因此,硅颗粒法是从鸟类陈旧标本骨骼组织提取DNA的理想方法,蛋白酶K法可用于鸟类陈旧标本羽毛的DNA提取.To find the better methods for extracting genomic DNA from old specimens of wetland bird, three methods, Proteinase K method, Chelex 1- 100 method, and Silica-based method,were used to extract genomic DNA from the feather, skin and bone tissues of the specimens. The obtained target fragments of 12S rDNA in the mtDNA were sequenced after the PCR. amplification of the DNA and then were compared with the Genbank data. The results show that the old specimens of wedand bird could be used for DNA extraction. Protein- ase K method and Silica-based method both can extract genomic DNA from the bone tissue of Egretta garzetta specimen and obtain 425 bp target fragments of 12S rDNA in the mtDNA（ the receive number of Genbank, AY766387 ）, and Proteinase K also can extract genomic DNA from the feather of Ciconia ciconia specimen and obtain 263 bp target fragments of 12S rDNA in the mt.DNA（ the receive number of Genbank, AY766388）. However, Chelex R 100 method cannot extract any genomic DNA from old specimens ofwedand bird. More DNA could be extracted by using Proteinase K method. But Proteinase K method needs more steps in the DNA purification, which increase the probability of external DNA pollution. Therefore, particular attentions should be paid to avoid the external pollution when using Proteinase K method, and the DNA extracted from the bone tissue has better to be purified. The silica-based method is a simple one for DNA extraction, which not only can decrease the pollution probability but also the DNA extracted from the bone tissue needn＇t to be purified, although the silicabased method extracted less DNA from the bone tissue than Proteinase K method and couldn＇t extract DNA from the feather. It is suggested that silica-based method is used for DNA extraction from the bone tissue of bird specimens and the Proteinase K method is used for DNA extraction from the feather.

关 键 词：DNA提取 鸟类标本 线粒体DNA 白鹭 白鹳 

分 类 号：Q959.7[生物学—动物学] S792.14[农业科学—林木遗传育种]