摘要
为建立一套从猪口腔中采集唾液并高效地从唾液中提取DNA的方法,选取约15日龄的仔猪和180日龄成年公猪各5头,分别用不同唾液采样工具来采集唾液样品并进行DNA提取,通过NanoDrop 2000和琼脂糖凝胶电泳检测DNA的浓度和纯度;同时以猪肌肉组织样品提取的DNA为参照,对比不同方法和不同日龄猪唾液提取的DNA质量差异,PCR扩增GAPDH基因以验证唾液DNA的可用性。结果显示:针对不同日龄的猪可以利用棉签和海绵采样工具进行唾液采集;建立的裂解法可以成功地从猪唾液中提取DNA,比人唾液DNA提取的碘化钾法质量更优;DNA纯度、浓度以及GAPDH基因扩增产物与肌肉组织提取的DNA效果相比趋近一致。提示:本文建立的猪唾液DNA的获取方法成功。
The purpose of this study was to establish a method for collecting saliva from the oral cavity of pigs and efficiently extracting DNA from these saliva samples.Saliva samples of five 15-day-old piglets and five 180-day-old boars were collected using different sampling tools.The DNA samples from these saliva samples were extracted,and their qualities were detected using NanoDrop 2000 and agarose gel electrophoresis.At the same time,DNA extracted from muscle samples was used as a reference for comparing the difference in the quality of saliva DNA samples originated from pigs at different ages or comparing the different extracting methods.Finally,the GAPDH gene was amplified by PCR in order to verify the availability of saliva DNA of pigs.Our study found that the saliva samples from pigs at different ages could be obtained using different saliva collection methods.Compared with the potassium iodide method,a common way for extracting human saliva DNA,lysis method can get better quality of DNA samples.Furthermore,the similar results of DNA purity,DNA concentration,and GAPDH amplification product were observed in the DNA samples extracted from the saliva and the muscle samples,indicating that the methods for both DNA collection and extraction from saliva of pigs were successfully developed in this study.
作者
李朝阳
罗武
杨凯
邵勇钢
翟曼君
董新星
韦伟
陈杰
张立凡
LI Chaoyang;LUO Wu;YANG Kai;SHAO Yonggang;ZHAI Manjun;DONG Xinxing;WEI Wei;CHEN Jie;ZHANG Lifan(College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China;College of Animal Science,Xinjiang Agricultural University,Urumqi 830052,China;College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China)
出处
《畜牧与兽医》
北大核心
2021年第6期7-11,共5页
Animal Husbandry & Veterinary Medicine
基金
中央高校基本科研业务费专项资金(KYYJ202008)
转基因生物新品种培育重点课题(2018ZX0800928B)
江苏省自然科学基金面上项目(BK20181319)。
