摘要
目前微量DNA的提取方法,不仅操作繁琐耗时,而且所需试剂价格昂贵。为了解决这个问题,本研究旨在开发一种快速、经济的微量DNA提取剂。用自制提取剂快速提取50个、100个、500个、5000个、50000个猪胎儿成纤维细胞的基因组DNA。经PCR扩增,电泳结果显示:自制提取剂可有效提取数量低至50个的猪胎儿成纤维细胞。分别采用自制提取剂、酚氯仿以及试剂盒3种方法提取猪单个囊胚基因组DNA,结果显示:酚氯仿方法提取的单个囊胚基因组DNA,经PCR扩增后,并没有检测到目的条带;而自制提取剂提取模板DNA,能直接扩增出清晰的目的条带,与试剂盒扩增效果相当。自制提取剂能够快速、有效地提取少量细胞和单个囊胚的基因组DNA,满足下游分子生物学研究需要,因此,本研究为微量样本基因组DNA提取提供帮助。
At present, the extraction method of small-scale DNA is not only complicated and time-consuming, but also expensive. The aim of this study was to develop a fast and economical micro DNA extractor agent to solve this problem. Genome DNA were quickly extracted by self-made extractor agent from 50, 100, 500, 5 000, 50 000 pig fetal fibroblasts, the results of PCR amplification showed that the self-made extractor agent could effectively extract even less 50 pig fetal fibroblasts. The genome DNA of pig single blastocyst was extracted by self-made extractor, phenol chloroform and kits, the results showed that the genomic DNA extracted from the single blastocyst by phenol chloroform did not detect the target band after PCR amplifying. However, the template DNA which extracted by self-made extractor could directly amplify clear target strips by PCR amplify directly, which has the same effect as the kit. The self-made extractor can extract the genome DNA rapidly and effectively from litter cells and single blastocyst to meet the needs of downstream molecular biology research. Therefore, this study provides help for genomic DNA extraction from micro samples.
作者
游小燕
范骁琳
何琦琳
刘雪芹
You Xiaoyan;Fan Xiaolin;He Qilin;Liu Xueqin(Chongqing Academy of Animal Science,Chongqing,402460;Key Laboratory of Pig Industry Sciences,Ministry of Agriculture,Chongqing,402460;Chongqing Key Laboratory of Pig Industry Sciences,Chongqing,402460)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2019年第9期4008-4012,共5页
Genomics and Applied Biology
基金
重庆市基本科研业务费项目(16419)资助
