big flowchart outlined - sea-phages · t ample e y er er es y esuspend hage hage with all edia er e...
TRANSCRIPT
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