Phage isolation, characterization, analysis and deposition

Benefit description

Bacteriophage Repository. Offers two types of repository: 1. Bacteriophage/host strain repository + information about each available on the website. 2. Hidden repository for users, only storage of phage/host strain samples. Includes annual viability testing and titration. 2. Bacteriophage Supply. Destinations: Research (public and private entities), compassionate use in hospitals (subject to approval by the competent authorities), and commercialization through companies (following the relevant agreements). 3. Host strain supply. These are intended for the propagation of bacteriophages to which they are sensitive. 4. Isolation of 1 bacteriophage against a bacterial species or strain. This will be performed from a sample provided or not by the user, and at least one phage will be isolated against a specific bacterial strain or species, sent by the user or from the collection itself. 5. Isolation of 5 bacteriophages against a bacterial species or strain. This will be carried out from a sample, whether provided by the user or not, and at least 5 phages will be isolated against a specific bacterial strain or species, either sent by the user or from the collection itself. 6. Propagation of bacteriophages in solid medium. It is essential that both the phage and the host bacteria be identified and sent as pure cultures. The final concentration of the suspension obtained may vary depending on the type of phage and its propagation efficiency on the host strain. 7. Propagation of bacteriophages in liquid media. It is essential that both the phage and the host bacteria be identified and sent in pure cultures. The service allows scaling up propagation in volumes of up to 2 liters. The final concentration of the suspension obtained can vary depending on the type of phage and its propagation efficiency on the host strain. 8. Titration of phage suspensions. Counting the number of viable phage particles per volume of the suspension in the host strain or in other strains sensitive to it. 9. Concentration of phage suspensions. Starting from a phage suspension, the number of particles per unit volume is increased, usually by PEG precipitation. 10. Cesium chloride gradient purification. Starting from a phage suspension, the number of particles per unit volume is increased, while simultaneously purifying them of any cellular debris, using a density gradient. 11. Prophage induction with mitomycin C. From lysogenic strains, prophages will be released by induction and isolated for subsequent characterization. 12. Phage DNA extraction (1 sample). DNA will be obtained from a suitably concentrated phage suspension, with a purity suitable for subsequent sequencing. 13. Phage DNA Extraction (10 samples). DNA will be obtained from 10 suitably concentrated phage suspensions, with a purity suitable for subsequent sequencing. 14. Phage DNA Sequencing. Once the DNA quality has been verified, it will be sent to an external company. 15. Sequence Assembly and Annotation. The sequenced fragments will be joined together to reconstruct the complete genome. Annotation is then performed, identifying genes, promoters, and regulatory elements. 16. Basic Analysis of Phage Genomic Sequences. Comparative analysis with the sequences of other phages to identify evolutionary and phylogenetic relationships. 17. Submission of Sequences to Databases. Deposit of genomic sequences in public or private repositories for storage, access, and analysis. These databases allow sharing genetic information with the scientific community. 18. Phagegrams. The resistance/sensitivity pattern of bacterial strains against one or more bacteriophages will be determined. The service offers the possibility