Big Flowchart Outlined - SEA-PHAGES · t ample e y er er es y esuspend hage hage with ALL edia er e...
Transcript of Big Flowchart Outlined - SEA-PHAGES · t ample e y er er es y esuspend hage hage with ALL edia er e...
Collect Environmental Sample
Collect Lysate from Plate with Highest Plaque Density
Spot Titer
Full Plate Titer
Prepare Webbed Plates
Plaque Assay
Pick a Plaque & Resuspend
Serially Dilute Sample with Phage
Extract Phage with SMALL
Volume of Media
Spin & Filter to Remove
Particulate & Bacteria
Test for Phage by Plaque Assay
Extract Phage with LARGE
Volume of Media
Spin & Filter to Remove
Particulate & Bacteria
Add Host Bacteria& Incubate
Spin to Remove Host Bacteria
Test for Phage by Spot Test
Collect Lysate from Webbed Plates
Spot Titer
Full Plate Titer
?Sample Contains Phage?
? Plaque Morphologies Consistent?
? Titer > 10 pfu/ml?9
ISOLATION PURIFICATION
Image by Electron Microscopy
MICROSCOPY
AMPLIFICATION
Compare Phages by Restriction Digest of DNA & Gel Electrophoresis
Extract Phage DNA
EXTRACTION
CHARACTERIZATION
Archive & Create phagesDB Entry