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AAT Bioquest

What are the methods for plasmid detection?

Posted January 20, 2023


Answer

There are several methods that can be used for plasmid detection. The 4 main methods include PCR, PFGE, optical mapping, and sequencing. PCR-based replicon typing is used to detect plasmids’ conserved replicon sites. Multiplex PCR can be expanded to target multiple replicon sites simultaneously. This is a quick and inexpensive method for plasmid detection. The downside is that detecting novel plasmid groups can be challenging. Optical mapping of plasmids involves stretching the plasmid DNA, which reveals the sequence of a plasmid. The downside of optical mapping is that it is unsuitable for detecting short plasmids that are less than 50 Kb. Pulsed field gel electrophoresis or PFGE helps in plasmid detection by applying an electrical field that changes direction periodically, which separates digested DNA on a gel matrix. The downside of this technique is that it can take several days to detect the number of plasmids and their size in an isolate. Sequencing-based plasmid detection uses high-throughput sequencing technologies that allow researchers to obtain all the information in plasmids. Plasmid and bacterial sequence data can be captured, extracted, and assembled using whole genome sequencing. Complete plasmid DNA sequencing is a powerful tool that allows researchers to obtain the full plasmid sequence in order to comprehensively analyze plasmids.

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