摘要
以冻鱼块、烤鱼片和鱼罐头3种主要水产加工品为研究对象,比较CTAB法、改良CTAB法、酚-氯仿法、SDS法以及5种试剂盒法等9种方法对3种水产加工品DNA的提取效果。通过比较DNA得率以及纯度,对各提取方法进行评价,并采用PCR方法对实验样品的真伪进行了初步鉴别。结果表明,9种提取方法对于3种水产加工品所提取DNA得率相差较大。对于冻鱼块采用深加工食品DNA提取试剂盒提取DNA的得率最高(1.223μgmg),A260/A280比值为2.0;对于烤鱼片采用SDS法提取得率最高(0.505μgmg),A260/A280比值为2.03;对于鱼罐头采用SDS法提取DNA得率最高为(0.594μgmg),A260/A280比值为1.93。所提取DNA可满足后续PCR鉴定实验的要求。本研究改进并筛选出水产加工品DNA提取的最佳方法,为后续水产加工品的原料真伪鉴别奠定了实验基础。
DNA was extracted from three different aquatic products including frozen fish block, grilled fish, and canned fish. Nine DNA extraction methods were compared, including CTAB precipitation method, improved CTAB precipitation method, Phenol - chloroform precipitation method, SDS precipitation method, and five genomic DNA purification kit. Quantity and quality of DNA was evaluated by the yield and the purity. Also, quality of the samples was evaluated by two couples of primers amplifying fragments of COI and Cytb. Results showed that DNA extraction from aquatic products by different processing methods could be employed with a specific DNA extraction method in each case. Best results were obtained with TIANGEN GMO food DNA extraction kit for frozen fish block in which the yield was 1. 223 μg/mg and the purity was 2.0 ; SDS method for grilled fish products the yield was 0. 505 μg/mg, and the purity was 2.03 ; SDS precipitation method for canned fish products the yield was 0. 594 μg/mg, amd the pu- rity was 1.93. The extracted DNA could meet the requirements of subsequent identification experiment. This study improved and screened the best method to extract DNA in aquatic products, and laid the experiment foundation for the subsequent identification of raw materials of processed aquatic products.
出处
《中国渔业质量与标准》
2013年第2期80-88,共9页
Chinese Fishery Quality and Standards
基金
科技支撑计划(2012BAD28B00)
水产行业标准项目(2013-407)
关键词
水产加工品
DNA提取
方法比较
真伪鉴别
aquatic products
DNA extracted methods
comparison
identification
