M. Dimitrakopoulou, Venia Stavrou, C. Kotsalou, A. Vantarakis
{"title":"煮沸萃取法与商业试剂盒分离食品样品细菌DNA的比较","authors":"M. Dimitrakopoulou, Venia Stavrou, C. Kotsalou, A. Vantarakis","doi":"10.26502/jfsnr.2642-11000057","DOIUrl":null,"url":null,"abstract":"Background: In recent years, the development of commercial Kits for DNA extraction, combined with the use of molecular technologies have been utilized for the understanding of microbial ecosystems. DNA extraction considered being a critical step for molecular techniques such as Polymerase Chain Reaction. Several different methods and commercial kits for this purpose are nowadays available. The aim of this study was to compare five commercially available kits and, a cheap, rapid, non-commercial method, the boiling method for isolating bacterial DNA from different food matrixes. Methods: Experimental protocol was applied on four different types of food: olives, roe, raisins and bacon. The evaluation of the results was performed first by using a spectrophotometer, by measuring purity and concentration of the extracted DNA, followed by electrophoresis where the integrity of the DNA was checked and finally by PCR-amplification. Results: Results revealed that boiling method is efficient, simple, cheap and suitable for PCR amplification for all these food types. Conclusion: Boiling method for bacterial DNA extraction from food should be further explored in order to be validated and established.","PeriodicalId":15858,"journal":{"name":"Journal of Food Science and Nutrition Research","volume":"24 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"11","resultStr":"{\"title\":\"Boiling Extraction Method VS Commercial Kits for Bacterial DNA Isolation from Food Samples\",\"authors\":\"M. Dimitrakopoulou, Venia Stavrou, C. Kotsalou, A. Vantarakis\",\"doi\":\"10.26502/jfsnr.2642-11000057\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Background: In recent years, the development of commercial Kits for DNA extraction, combined with the use of molecular technologies have been utilized for the understanding of microbial ecosystems. DNA extraction considered being a critical step for molecular techniques such as Polymerase Chain Reaction. Several different methods and commercial kits for this purpose are nowadays available. The aim of this study was to compare five commercially available kits and, a cheap, rapid, non-commercial method, the boiling method for isolating bacterial DNA from different food matrixes. Methods: Experimental protocol was applied on four different types of food: olives, roe, raisins and bacon. The evaluation of the results was performed first by using a spectrophotometer, by measuring purity and concentration of the extracted DNA, followed by electrophoresis where the integrity of the DNA was checked and finally by PCR-amplification. Results: Results revealed that boiling method is efficient, simple, cheap and suitable for PCR amplification for all these food types. Conclusion: Boiling method for bacterial DNA extraction from food should be further explored in order to be validated and established.\",\"PeriodicalId\":15858,\"journal\":{\"name\":\"Journal of Food Science and Nutrition Research\",\"volume\":\"24 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"11\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Food Science and Nutrition Research\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.26502/jfsnr.2642-11000057\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Food Science and Nutrition Research","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.26502/jfsnr.2642-11000057","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Boiling Extraction Method VS Commercial Kits for Bacterial DNA Isolation from Food Samples
Background: In recent years, the development of commercial Kits for DNA extraction, combined with the use of molecular technologies have been utilized for the understanding of microbial ecosystems. DNA extraction considered being a critical step for molecular techniques such as Polymerase Chain Reaction. Several different methods and commercial kits for this purpose are nowadays available. The aim of this study was to compare five commercially available kits and, a cheap, rapid, non-commercial method, the boiling method for isolating bacterial DNA from different food matrixes. Methods: Experimental protocol was applied on four different types of food: olives, roe, raisins and bacon. The evaluation of the results was performed first by using a spectrophotometer, by measuring purity and concentration of the extracted DNA, followed by electrophoresis where the integrity of the DNA was checked and finally by PCR-amplification. Results: Results revealed that boiling method is efficient, simple, cheap and suitable for PCR amplification for all these food types. Conclusion: Boiling method for bacterial DNA extraction from food should be further explored in order to be validated and established.