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Phage Plaque Test for Counting Viruses
Description
Bacteriophages, or phages, are viruses that infect prokaryotic cells. The plaque assay is a classic way to count these viruses by measuring viral titer, which is the amount of virus in a sample. It was first described in 1952 and is still one of the most reliable methods for determining viral titer.
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Transcript
噬菌体,也称为噬菌体(phages),是一类特异性感染细菌的病毒,我们可以通过一种称为噬菌斑实验(plaque assay)的工具来确认其存在并进行定量。噬菌体通过首先附着在细菌细胞壁上并将其遗传物质注入细胞内,从而感染敏感宿主。随后,它们劫持细胞的生物合成机制以复制自身DNA,并产生大量子代噬菌体颗粒,最终通过裂解宿主细胞并杀死该细胞而释放出来。
这种裂解活性是广泛使用的噬菌体计数技术——噬菌斑测定法(又称双层琼脂法)的基础。首先,在含有低浓度琼脂的熔融营养肉汤中制备噬菌体混合液。混合物中使用的所有细菌应处于对数生长期,保持活性并正在进行分裂,以确保大部分细菌具有活性,能够在噬菌斑周围形成致密的菌苔。随后,将该熔融的细菌-噬菌体-琼脂混合物铺展于已凝固在培养皿中的较坚固、...
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