摘要
目的为了探索一种既能使野外采集的粪便样品DNA保存完好,又方便带回实验室用于分子水平研究的粪便保存方法。方法本研究通过将普氏原羚的粪便在野外分别采取60℃干燥后低温保存,75％乙醇保存,100％乙醇保存,直接低温(保温箱中加生物冰袋)保存,在1个月内、5个月后、8个月后对粪便DNA进行抽提,并将提取的DNA作为模板进行PCR扩增,基因克隆测序。结果短期内均可得到高质量的DNA,DNA大小在23kb左右,电泳带型整齐,无降解,线粒体的扩增结果也完全一致,而用100％乙醇保存粪便DNA的时间更为长久。结论不同的保存方法提取动物基因组效果不一样。
Objective To find a method that it is facilitated to preserve the faeces in the field collection, and that it is also easy to carry back to the research laboratory for molecular level of faeces DNA samples. Methods Respectively, with the low temperature preservation of 60 degrees dry, the preservation of 75% ethanol, the preservation of 100% ethanol and direct low temperature（ in the attemperator adding the biological ice bag） , were to preserve the faeces from Przewalski＇s gazelle in the field of collecting. The DNA was extracted from faeces samples in one month, five months later and eight months later, respectively. The extraction of DNA as a template was used for PCR, gene cloning and sequencing. Results The results showed that DNA templates were extracted successfully from the faeces in a short time, The DNA molecular weight is about 23kb. The electrophoresis belts were tidy, without degradation. The expansion results of mitochondria are completely consistent. With the 100% ethanol, the DNA samples will be preserved for a longer time. Conclusions The different method preservation will have differently extraction results.
出处
《国际遗传学杂志》
CAS
2006年第5期341-345,367,共6页
International Journal of Genetics
基金
国家自然科学基金项目(No.30370217)
濒危动物保护技术研究项目(No.2001DIB10058)
关键词
粪便保存方法
DNA提取
普氏原羚
Preservation methods of faeces
Faecal DNA extraction
Przewalski＇s gazelle
