Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include 1 streak-plating bacterial cultures to isolate single colonies, 2 pour-plating and 3 spread-plating to enumerate viable bacterial colonies, 4 soft agar overlays to isolate phage and enumerate plaques, and 5 replica-plating to transfer cells from one plate to another in an identical spatial pattern.
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Aseptic Laboratory Techniques: Plating Methods
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