摘要
细菌基因组DNA的提取质量直接影响以PCR为基础的快速检测结果,影响检测速度、灵敏度、检测限度等指标。其中菌体的培养浓度是重要的一个环节,过浓或浓度太低都会对后续的PCR操作带来干扰。为简单方便的测定菌体培养液的浓度,用麦氏比浊仪测定培养液中的菌体浓度,同时用平板计数法进行验证,结果表明麦氏比浊仪可达到测定目的,制定了相关菌种的标准曲线,可快速准确的测定菌液浓度。同时通过测定提取到的基因组含量,验证了与菌体浓度的相关性。另外也对比了不同的基因组提取方法的提取效率并评价,选择了最适合本试验的方法三:改良Universal Genomic DNA Extraction Lit Ver.3.0方法。
Bacterial genomic DNA extraction quality directly affect the rapid test results based on PCR,including the detection speed,sensitivity,detection limit and other indicators.The concentration of cell culture is very important,too thick or too low concentrations will bring interference to the subsequent operation of PCR.In order to get the concentration of cell culture simply and conveniently,the Maxwell nephelometer were used to determine the cell concentration in the supernatant,at the same time validated the data using the plate count method.It showed that Maxwell nephelometer could match the determination purpose and measure the concentration of bacteria quickly and accurately.The standard curve of bacteria concentration were made,compared with the genome content,they have relevance.Different methods of extracting genome were compared to determine the extraction efficiency,at last the third：modified Universal Genomic DNA Extraction Lit Ver.3.0method was selected.
出处
《食品研究与开发》
CAS
北大核心
2016年第4期172-175,共4页
Food Research and Development
关键词
基因组浓度
PCR
标准曲线
concentration of bacteria genome
polymerase chain reaction（PCR）
standard curve
