奶牛瘤胃胃液微生物总DNA的提取和纯化

Extration and purification of DNA from evironmental rumen samples

本研究建立了从奶牛瘤胃液中提取混合微生物总DNA的方法,对现有的方法进行改进使其适合于瘤胃混合微生物总DNA的提取。粗提后的瘤胃DNA可以直接扩增出16SrDNA。提取效率为每毫升瘤胃液的DNA提取量为0.1～0.3μg。通过SiO2回收进行纯化,纯化回收率达到34%～50%;用cleanup试剂盒纯化,纯化率可达到85%以上。经过纯化后的DNA可进行其它的分子生物学操作。

A method has been developed for extracting and purifying genomic DNA from environmental rumen samples. On the basis of this method, an environmental sample is treated first by feezing/ thawing and subsequently by SDS/ proteinase K based DNA extraction. The yields of purified DNA from three samples used in this study ranged from 0.1 to 0.3μg /mL of fluid sample. The crud mixed genomic DNA can be used as templet for PCR. The crud mixed genomic DNA of the environmental samples was purified by SiO2. method and clean-up kit (Promega).Their rate of purification is 34%～50% and 85% respectively. The purified DNA can be used in molecular experiment.