After years of exploration and expansion, Creative Biolabs has become a recognized leading expert in the field of phage research. We are always willing to share cutting-edge technologies and information about phage research with you. Additionally, we offer comprehensive phage analysis services, from phage isolation to final result analysis.
Bacteriophages are well known to be simple organisms consisting of a protein capsid and nucleic acid. Notably, nucleic acids can be DNA or RNA and can be double-stranded or single-stranded. A solid understanding of phages contributes to elucidating the mechanisms of molecular biology and evolutionary genetics. Furthermore, bacteriophages have attracted increasing attention due to their important role in alleviating the current medical challenges posed by antibiotic resistance. Phages are typically isolated and subsequently characterized to obtain phages with the desired properties.
Phage isolation is generally fast, simple, and inexpensive compared to other antimicrobial agents. There are many ways to obtain target phages, but they are all based on similar patterns. A commonly used method is described as an enrichment procedure. The phage-containing sample is first mixed with host bacteria and the mixture is incubated for a period of time, usually overnight. The remaining bacteria were then removed from the cultures by centrifugation and filtration, and the filtrate was assayed for the presence of phage. Any phages identified were then characterized to determine if they possessed the properties required for phage therapy.
Fig.1 Methodology for phage isolation. (Runa, 2021)
Various methods are available for phage purification. A combination of low-speed centrifugation and membrane filtration is the most basic method for purifying phage lysates. Furthermore, the most important phage purification method for small-scale preparations is high-speed centrifugation in the cesium chloride gradient. Notably, based on the fact that bacteriophage and endotoxin accumulate in different solvents, endotoxin can be removed by extraction with organic solvents. Currently, anion exchange chromatography is emerging as a new alternative.
Phages are considered one of the attractive alternatives to deal with the growing population of antibiotic-resistant bacteria. Phage amplification is a critical procedure as high titer preparations are beneficial in studies. Commonly used methods of phage amplification include the plate wash method, pooled plaques method, agar culture method, and two-stage culture method. It is worth noting that there is no universal phage amplification method, and it is vital to establish the most suitable method for each phage.
Fig.2 The schematic representation of the phage amplification assay methodology. (Beinhauerova, 2021)
Whole-genome sequencing contributes to a better understanding of bacteriophages, which requires high-quality phage DNA. In general, many methods of phage DNA extraction for genomics analysis are complex and time-consuming. Therefore, it is very necessary to develop fast and cost-effective methods. At Creative Biolabs, several efficient methods are available. In our assay, we prepared highly concentrated phage lysates using acetone precipitation, followed by extraction using commercial kits or potassium iodide. The quality of the extracted DNA was analyzed by agarose gel electrophoresis and DNA analysis. Finally, the extracted DNA was subjected to restriction digestion and next-generation sequencing.
DNA sequencing technology provides a powerful tool for the analysis of phage genomes. Genome sequencing and characterization of isolated phages can indicate the presence of virulence or lysogenic genes, which provides the basis for excluding non-usable phages. Currently, many computational tools play an important role in phage identification or classification.
With our proven capabilities and advanced technology, we are confident in providing high-quality services for phage analysis and characterization, such as phage isolation, purification, amplification, and DNA extraction. We offer many flexible options, and you can always find one that better matches your phage discovery project. Please contact us for more details.
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