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8.4K Views
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06:12 min
October 25th, 2018
DOI :
10.3791/58612-v
Chapters
0:04
Title
0:30
Sample Preparation
1:39
Immunomagnetic Separation (IMS) and Multiple Displacement Amplification (MDA)
3:37
Real-time PCR and Library Preparation for Quasimetagenomic Sequencing
4:52
Results: RT-PCR Amplifies DNA from Two Brands of Salmonella Contaminated Chicken Breast
5:19
Conclusion
Transcript
该方法可用于检测食品样品中的沙门氏菌,并通过基因指纹识别病原体至菌株水平。该技术的主要优点是,它结合了沙门氏菌检测和亚分到一个工作流程,并大大缩短了从沙门氏菌污染食品样品到病原体指纹的周转时间。首先,无菌地将 25 克食物样品放入无菌实验室搅拌袋中,并配有内置过滤器,或者可选地准备环境拭子,用浓缩汤对海绵进行无菌润湿,将其拖过预先确定的表面,然后放入搅拌袋中。
使用实验室搅拌机,将每个样品与 225 毫升 RV 浓缩汤彻底混合。然后在42摄氏度下孵育混合物4至21小时。在此之后
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Summary
在这里, 我们提出了一个协议, 准备从食物和环境微生物群 DNA 样本, 以协同检测和子类型沙门氏菌通过 quasimetagenomic 测序。结合使用培养富集、免疫磁分离 (IMS) 和多重置换扩增 (MDA), 可有效地将沙门氏菌基因组 DNA 从食物和环境样本中浓缩。
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