烤鳕鱼片中基因组DNA提取条件优化及其应用  

Optimization on Genome DNA Extraction Conditions of Roasted Cod Fillets and Its Application

作　　者：刘珊珊 李岩 陈光 赵冬梅 张利伟 朱娜 LIU Shanshan;LI Yan;CHEN Guang;ZHAO Dongmei;ZHANG Liwei;ZHU Na(Qinhuangdao Food and Drug Inspection Center,Qinhuangdao 066000;Hebei Provincial Medical Products Professionalization Inspector Corps,Shijiazhuang 050000)

机构地区：[1]秦皇岛市食品药品检验中心,秦皇岛066000 [2]河北省药品职业化检查员总队,石家庄050000

出　　处：《食品工业》2022年第3期41-44,共4页The Food Industry

基　　金：河北省市场监督管理局科研计划项目(2020ZD35)。

摘　　要：为了对烤鳕鱼片中的鳕鱼成分进行实时荧光PCR检测,将烤鳕鱼片基因组DNA的提取条件进行响应面法优化研究,以CTAB为裂解液,考察取样量、蛋白酶量、裂解温度等因素对提取质量的影响。每个因素的最佳水平由单因素试验确定。选择Box-Behnken方案进行设计并试验,以最终DNA浓度为响应值,利用响应面法对参数进行优化,最终得到烤鳕鱼片中基因组DNA的最佳提取条件:取样量0.2 g、加酶量45μL、裂解温度65℃。在该条件下进行试验,提取的基因组DNA实际浓度为627.0±13.0 ng/μL,与模型预计结果637.5 ng/μL的相对误差为1.66%,误差较小,适用于烤鱼片中基因组DNA的提取。利用优化方法对6种市售秦皇岛烤鳕鱼片进行DNA提取并进行鳕鱼成分的实时荧光PCR检测,所得结果均检出鳕鱼成分。In order to detect cod ingredients in roasted cod fillets by real-time PCR,response surface methodology was used to optimize the DNA extraction conditions of roasted cod fillets.With CTAB as lysis solution,the effects of sampling weight,volume of proteinase and pyrolysis temperature on the DNA concentration were investigated.The optimum level of each factor was determined by single factor test.The Box-Behnken scheme was selected for design and test.With the final DNA concentration as the response value,the parameters were optimized by response surface methodology.The best optimum extraction conditions of genome DNA in roasted cod fillets were as follows:sampling weight 0.2 g,volume of proteinase 45μL and pyrolysis temperature 65℃.Under these conditions,the final concentration of DNA from roasted cod fillets was 627.0±13.0 ng/μL.The relative error with the model prediction result of 637.5 ng/μL was 1.66%.The error was small,so the model was suitable for the DNA extraction conditions of roasted cod fillets.The optimized method was used to extract DNA from 6 kinds of roasted cod fillets sold in Qinhuangdao,and the cod ingredients were detected by real-time fluorescence PCR.

关 键 词：DNA提取 响应面法 烤鱼片 实时荧光PCR 鳕鱼 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]