摘要
为了筛选出一种快速、高效、廉价提取猪肺炎支原体模板DNA的方法,本研究分别对试剂盒法、双蒸水煮法、酚-氯仿法进行比较,从中选出最佳提取方法。结果显示:这3种方法均适合由菌液中提取基因组DNA;水煮法不能由组织中提取出基因组DNA,试剂盒法可由组织中提取出基因组DNA且得到的目的DNA较纯,但每次提取量少;酚-氯仿法能大批量提取组织基因组DNA,但操作繁琐、易污染且稍有杂带。针对不同的模板,选择合适的DNA制备方法,有利于做出准确检测、提高检测效率,为疾病的早期诊断提供保障。
In order to filter out a fast,efficient,inexpensive method to extract the Mycoplasma hyopneumoniae DNA tempiate.In this study we compared Tissue DNA Kit,double distilled boiled method and phenol-chloroform method to choose a best way to extract Mhp DNA.The results showed that the three methods were suitable for the extraction of genomic DNA of the bacterium,however,double distilled boiling method could not extracted genomic DNA from the tiusse,the Tissue DNA Kit could extract out of genomic DNA purely from the tissue,but got a small quantity of DNA each time,phenol-chloroform method could extract large DNA of the tissue,but complicated to operate,vulnerable to pollution,and with a little impurity bands.Acorrding to the different templates,select the appropriate DNA preparation method which is beneficial to make the detection accurated,meanwhile improve the detection efficiency,to provide protection for the early disease detection.
出处
《中国农学通报》
CSCD
2013年第29期52-56,共5页
Chinese Agricultural Science Bulletin
基金
江苏省自主创新项目"新型猪支原体肺炎活疫苗研制"(cx(11)4042)
关键词
猪肺炎支原体
猪支原体肺炎
基因组DNA
模板制备
PCR检测
Mycoplamsa hyopneumoniae
Mycoplasmal pneumonia of swine,genomic DNA
template preparation
PCR detection
