We present a modified alkaline lysis method for purification of plasmid DNA (pDNA) from bacterial extract using fractional precipitation with isopropanol (FPI). This method includes two successive precipitations with 0.33 and 0.36 volumes of isopropanol and separates pDNA from total RNA and most of the lipopolysaccharides. Using different quality control tests, we demonstrate that plasmids purified with FPI show superior quality compared to plasmids prepared with commercial kits based on spin-column chromatography.
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http://dx.doi.org/10.2144/btn-2021-0018 | DOI Listing |
NAR Genom Bioinform
March 2025
Departments of Medicine and Pediatrics, Division of Infectious Diseases and Global Health, University of California San Francisco School of Medicine, 550 16th Street, 4th Floor Mission Hall, San Francisco, CA, 94158, USA.
Whole genome sequencing (WGS) is pivotal for the molecular characterization of ()-the leading bacterial cause of sexually transmitted infections and infectious blindness worldwide. WGS can inform epidemiologic, public health and outbreak investigations of these human-restricted pathogens. However, challenges persist in generating high-quality genomes for downstream analyses given its obligate intracellular nature and difficulty with propagation.
View Article and Find Full Text PDFPLoS One
January 2025
Produce Safety and Microbiology Unit, Western Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Albany, California, United States of America.
Non-typhoidal Salmonella enterica is a leading cause of gastrointestinal illnesses in the United States. Among the 2,600 different S. enterica serovars, Infantis has been significantly linked to human illnesses and is frequently recovered from broilers and chicken parts in the U.
View Article and Find Full Text PDFSci Rep
January 2025
Microbiology Division, Defence Research and Developmental Establishment, Jhansi Road, Gwalior, 474002, India.
Yersinia pestis, a Gram-negative bacterium is the causative agent of the fatal communicable disease plague. The disease had a profound impact on human history. Plague bacteria are usually transmitted to humans through the bite of an infected rat flea.
View Article and Find Full Text PDFBMC Vet Res
January 2025
College of Life Science and Engineering, Foshan University, Foshan, Guangdong, 528231, China.
Background: Pseudorabies virus (PRV), porcine parvovirus (PPV) and porcine circovirus 3 (PCV3) are common in swine farms in China. Single infection or co-infection with PRV, PPV and/or PCV3 was difficult to distinguish between their clinical symptoms and pathological changes. Therefore, a quick and accurate detection method is needed for epidemiological surveillance, disease management, import and export control.
View Article and Find Full Text PDFPathogens
November 2024
College of Animal Science and Technology, Guangxi University, Nanning 530005, China.
Porcine astrovirus (PoAstV), porcine sapovirus (PoSaV), porcine norovirus (PoNoV), and porcine rotavirus A (PoRVA) are newly discovered important porcine diarrhea viruses with a wide range of hosts and zoonotic potential, and their co-infections are often found in pig herds. In this study, the specific primers and probes were designed targeting the ORF1 gene of PoAstV, PoSaV, and PoNoV, and the VP6 gene of PoRVA. The recombinant standard plasmids were constructed, the reaction conditions (concentration of primers and probes, annealing temperature, and reaction cycle) were optimized, and the specificity, sensitivity, and reproducibility were analyzed to establish a quadruplex real-time quantitative RT-PCR (RT-qPCR) assay for the detection of these four diarrheal viruses.
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