浙江农业学报 ›› 2019, Vol. 31 ›› Issue (9): 1471-1477.DOI: 10.3969/j.issn.1004-1524.2019.09.10

• 动物科学 • 上一篇    下一篇

几种动物生鲜肌肉组织样品DNA提取方法的比较研究

纪艺1,2, 姜媛媛1,2, 汪小福2, 徐晓丽2, 徐俊锋2, 李玥莹1,*, 陈笑芸2,*   

  1. 1.沈阳师范大学 生命科学学院,辽宁 沈阳 110034;
    2.浙江省农业科学院 农产品质量标准研究所,省部共建农产品质量安全国家重点实验室(筹),浙江 杭州 310021
  • 收稿日期:2019-07-09 出版日期:2019-09-25 发布日期:2019-10-11
  • 通讯作者: *李玥莹,E-mail:yueyinglicn@163.com;陈笑芸,E-mail: xiaoyunchen_2016@163.com
  • 作者简介:纪艺(1995—),女,辽宁铁岭人,满族,硕士研究生,专业为生物化学与分子生物学。E-mail: jymemory12138@163.com

Comparative study of several DNA extraction methods for raw animal muscle tissues

JI Yi1,2, JIANG Yuanyuan1,2, WANG Xiaofu2, XU Xiaoli2, XU Junfeng2, LI Yueying1,*, Chen Xiaoyun2,*   

  1. 1. College of Life Science, Shenyang Normal University, Shenyang 110034, China;
    2.State Key Laboratory for Quality and Safety of Agro-Products(in Prepared), Institute of Quality and Standard for Agro-Products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
  • Received:2019-07-09 Online:2019-09-25 Published:2019-10-11

摘要: 比较不同提取原理的试剂盒对动物生鲜肌肉组织DNA提取效果,为日常肉制品掺假检测与监测选择合适的DNA提取方法提供参考。以生鲜牛肉、羊肉、猪肉、鸡肉和鸭肉为试验材料,采用改良CTAB法、蛋白酶K法、SDS法和快速萃取法等原理的8种不同方法来提取组织样品DNA,并对提取DNA的完整性、浓度、纯度、试验耗时和后续实时荧光定量PCR检测等指标进行评价。结果表明,8种提取方法均可用于常见动物源性肌肉DNA的提取。与改良CTAB法相比,原理为蛋白酶K法、SDS法和快速萃取法的试剂盒避免了有机试剂的使用,且操作耗时短。实际操作中,可优先选择基于蛋白酶K法和SDS法的试剂盒进行DNA提取,其提取的DNA纯度和总量高,盐残留少,对实时荧光定量PCR的抑制少,满足肉制品掺假检测与监测的核酸提取需求。

关键词: DNA提取, 实时荧光定量PCR, 肌肉组织, 评估

Abstract: DNA extraction methods with different principles for raw animal muscle tissue were conducted and compared to define the most suitable extracting method for the daily work of detecting as well as monitoring the adulteration in meat and meat products. The eight DNA extracting methods with various principles included the traditional CTAB-, proteinase K-, SDS-and rapid extraction-based methods, and the DNA extraction were conducted on raw beef, pork, lamb, chicken and duck. Moreover, the extracted DNA was evaluated for the integrity, yield, purity, time-consuming and inhibition effects on the real-time fluorescence quantitative PCR. As a result, all eight methods could be utilized to achieve the DNA extraction from the muscle tissues of common varieties of meat. Generally, all the commercial kits that based on the proteinase K, SDS or rapid extraction principle avoided organic solvents and exhibited a time-saving character when compared with the traditional CTAB-based method. Moreover, regarding to the higher DNA yield and purity as well as the less time-consuming and little fluorescence inhibition in real-time PCR, it gave priority to choose the commercial kits with proteinase K-or SDS-based method to extract DNA. Accordingly, these commercial kits could meet the requirement of the DNA extraction for detecting or monitoring the adulteration in raw muscle tissues of common varieties of meat.

Key words: DNA extraction, real-time PCR, muscle tissue, evaluation

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