For Direct Isolation For Enriched Isolation Extract phage w enrichment brothmedia Filter to remove soil bacteria Test via Plaque Assay Confirm w Spot Test Pick a Plaque OR Serial Dilutions ID: 1038110
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1. Collect Environmental SamplesFor Direct IsolationFor Enriched IsolationExtract phagew/ enrichment broth/mediaFilter to remove soil bacteriaTest via Plaque AssayConfirm w/ Spot Test Pick a PlaqueORSerial Dilutions1-2 hrs1-2 hrsFilter to remove soil bacteriaAssay via Spot Test10-110-210-310-410-510-110-210-310-410-5Plaque Assay for PurificationPick a Plaque10-110-210-310-410-5100Serial DilutionsPlaque Assay10-110-210-310-410-5100Are your plaque morphologies consistent?YesNOFlood bestWebbed plate Collect Small Volume LysateSpot Titer10-510-110-210-310-410-6Full Plate Titer10-510-410-6Make Webbed Plates Collect Large Volume LysateTransmission Electron MicroscopyExtract Phage DNARestriction Enzyme DigestGel electrophoresisSelect DNA for sequencing!ArchivingandPhagesDBIf your titer > 5x 109Spot Titer10-510-110-210-310-410-6Full Plate Titer10-510-410-6FloodIf your titer < 5x 109Extract phagew/ enrichment broth/mediaSeed w/ host bacteriaFilter to remove bacteria48 hrs