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A simple and rapid technique of template preparation for PCR

Frontiers in Microbiology, ISSN: 1664-302X, Vol: 13, Page: 1024827
2022
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Researchers from Chinese Academy of Agricultural Sciences Discuss Research in Microbiology (A simple and rapid technique of template preparation for PCR)

2022 NOV 24 (NewsRx) -- By a News Reporter-Staff News Editor at NewsRx Life Science Daily -- Current study results on microbiology have been published.

Article Description

Many techniques have been developed for extracting DNA, but most are often complex, time-consuming, and/or expensive. In this study, we describe a simple, rapid and cost-effective technique for preparing DNA template for PCR. This technique involves 0.1 M potassium hydroxide treatment at 100°C for 10 min followed by centrifugation. The suspended centrifuged sediments were shown as excellent templates for PCR. Templates prepared using this technique worked for diverse microorganisms belonging to bacteria, fungi and oomycetes and their amplification efficiencies were comparable to/better than those prepared using common but relatively more complex, time-consuming, and/or expensive methods, including commercial DNA extraction kits. Furthermore, this technology is suitable for high-throughput batch processing and for amplifications of long DNA fragments. Flow cytometry and scanning electronic microscopy analyzes showed that this technique generated primarily damaged cells and cell-bound DNA, not free naked DNA. This technique provides a great convenience for simple PCR template preparation.

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