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食品研究与开发:2018,39(2):177-180
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采用碱裂解法提取DNA鉴定细菌和酵母菌
迟原龙1,2,唐垚1,金璐1,张凤1,姚开1,何强1
(1.四川大学轻纺与食品学院,四川成都610065;2.成都市食品药品检验研究院,四川成都610041)
Identification of Bacteria and Yeasts by Using An Alkaline Lysis Method for DNA Extraction
CHI Yuan-long1,2,TANG Yao1,JIN Lu1,ZHANG Feng1,YAO Kai1,HE Qiang1
(1.College of Light Industry and Food Engineering,Sichuan University,Chengdu 610065,Sichuan,China;2.Chengdu Food and Drug Administration Research Institute,Chengdu 610041,Sichuan,China)
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投稿时间:2017-06-09    
中文摘要: 采用碱裂解法对分离自四川泡菜的8株细菌和4株酵母菌进行DNA提取,对菌株DNA提取物浓度和纯度、PCR扩增产物质量、核苷酸测序和菌株鉴定结果进行分析,并对该方法的测试成本和耗时进行估算。结果表明,采用碱裂解法提取菌株DNA的浓度均大于40 ng/μL,A260/A280大于1.9,表明该方法下提取菌株DNA的质量较好。12株泡菜菌株被鉴定至种水平,且鉴定结果与试剂盒法一致,表现出较高的测试准确性。该方法的测试成本估算为50元/100次,测试耗时为50 min,明显优于目前普遍使用的试剂盒法。
中文关键词: 碱裂解法  DNA提取  四川泡菜  菌种鉴定
Abstract:Genomic DNA of 8 strains of bacteria and 4 strains yeasts,isolated from Sichuan pickles,was extracted by using an alkaline lysis method.Concentration and purity of strain DNA extracts,quality of PCR amplified products,results of nucleotide sequencing and strain identification were studied.Furthermore,the cost and time-consuming of this method were evaluated.Results showed that the DNA concentration and(A260/A280)of the extracts were all>40 ng/μL and>1.9,respectively.It suggested good quality of the DNA extracts.12 strains isolated from Sichuan pickles were identified into species level,and their results were consistent with that obtained from test kit assays,which indicates a high accuracy of the method.The cost and time-consuming of the method were evaluated as fifty yuan per 100 times and 50 min,respectively.And so,it was much better than the test kit assays used currently.
文章编号:201802032     中图分类号:    文献标志码:
基金项目:国家自然科学基金(31601442);成都市产业集群协同创新重大项目(2016-XT00-00031-NC)
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