Bacteriophages are the most abundant biological entity on earth, with an estimated number of 1031.  Bacteriophages specifically kill bacteria.  Phages that infect M.smegmatis could potentially be used as an alternative to antibiotics.https://digitalcommons.mtech.edu/urp_aug_2018/1012/thumbnail.jp

Pedulla, Marisa L

Sparks, Hannah E

English

Digital Commons @ Montana Tech

Montana Tech LibraryDigital Commons @ Montana Tech2018 Undergraduate Research Other Undergraduate ResearchSummer 2018Curation and Characterization of a Collection ofMycobacteriophagesHannah E. SparksMontana TechMarisa L. PedullaMontana TechFollow this and additional works at: https://digitalcommons.mtech.edu/urp_aug_2018This Book is brought to you for free and open access by the Other Undergraduate Research at Digital Commons @ Montana Tech. It has been acceptedfor inclusion in 2018 Undergraduate Research by an authorized administrator of Digital Commons @ Montana Tech. For more information, pleasecontact sjuskiewicz@mtech.edu.Recommended CitationSparks, Hannah E. and Pedulla, Marisa L., "Curation and Characterization of a Collection of Mycobacteriophages" (2018). 2018Undergraduate Research. 13.https://digitalcommons.mtech.edu/urp_aug_2018/13Curation and Characterization of a Collection of MycobacteriophagesHannah E. Sparks & Dr. Marisa L. PedullaBacteriophage BackgroundMethodsConclusionsAcknowledgementsResultsGemmarlo Paolini Prickles MoonbeamTomaszewski TRexWhatsapiecost Rope• Bacteriophages are the most abundant biological entity on earth, with an estimated number of 1031.• Bacteriophages specifically infect and kill bacteria. • Phages that infect M.smegmatis could potentially be used as an alternative to antibiotics. • Isolation of single plaques from phages stored at various purification stages at Montana Tech. • Creation of a high titer sample.• DNA Extraction• Restriction Enzyme Digests & Gel Electrophoresis• Send sample for sequence analysis at the University of Montana• Polymerase Chain Reaction (PCR) for cluster analysis • Update Phage Information• Archive samples at the University of Pittsburgh• Upload information to phagesdb• Update spreadsheet of Montana Tech’s phages• Transmission Electron Microscopy. • Transmission electron microscopy indicated Siphoviridae morphology for all eight phages. • Restriction enzyme digests indicated multiple cut sites for all phage genomes, giving each phage a unique fingerprint. • Updating information of phages is important for future research. Future Work• Annotate genomic sequences when they are returned. • Repeat methods for more phages stored at Montana Tech. • Further discovery through bioinformatics or wet lab procedures. • “This work was supported by Montana Tech’s Summer Undergraduate Research Fellowship (SURF). We thank…• Student Researchers, Tavi Wise & Erin Fogarty • University of Missoula & Dr. Jim Driver, EMTrix• Dr. Joel Graff for resources utilized from his lab. • HHMI, SEA-phages, University of Pittsburgh• This research was supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number P20GM103474. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.”PCR Cluster AnalysisRestriction Enzyme DigestPhage NameConcentration from DNA extraction(ng/uL)Lysate Concentration (pfu/mL)Gemmarlo 14.2 3.7 x 1010Moonbeam 55.8 1.67 x 1010Paolini 34.3 1.67 x 1010Prickles 31.2 6.67 x 109Solarflare 72.9 In ProgressTomaszewsky 65.7 2.33 x 1010TRex 40.4 1.67 x 109Whatsapiecost 13.4 6.0 x 1010Phage Name Putative ClustersWhatsapiecost A1, A2, A6, B1, F2, KPrickles DMoonbeam B1, B3, D, I1/I2, A1, F2, ISolarflareA1, A2, B1, B3, D, E, F1, F2, G, I1/ I2, K, B4, C2Paolini A2Tomaszewsky A5, A6, B1, D, E, F1, F2, G, H, I1/I2, A1, B2, B3,E, IGemmarlo A2, A4, D, B3, E, I1/I2, K, GGL, A1, B1, B2, F2, F1, G, H2, ITRexA1, A2, A3, A4, A5E, F2, G, B3, C2 

Curation and Characterization of a Collection of Mycobacteriophages

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Curation and Characterization of a Collection of Mycobacteriophages

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