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The streaked plate is incubated at 37☌ for 24 hours. Note: Bi-plate inoculation of samples from sterile sites is often done in diagnostic laboratories to save time and space. Returning to the area you just streaked (area 3), extend the streaks into the center fourth of the plate (area 4). Flame the loop again and allow it to cool.Returning to the area you just streaked (area 2), extend the streaks into the third quarter of the plate (area 3). Returning to the edge of area 1 that you just streaked, extend the streaks into the second quarter of the plate (area 2).

Immediately streak the inoculating loop gently over a quarter of the plate using a back-and-forth motion (see area 1 in the figure above).

Pick an isolated colony from the agar plate culture and spread it over the first quadrant (approximately 1/4 of the plate) using close parallel streaks.

Sterilize the inoculating loop in the bunsen burner by putting the loop into the flame until it is red hot.

Quadrant Streaking for isolation into pure culture Procedure Usually, by the third or fourth quadrant, only a few organisms are transferred, giving discrete colony-forming units (CFUs). There are four basic types of streaking methods Īs the original sample is diluted by streaking it over successive quadrants, the number of organisms decreases. Many different streaking patterns can be used to separate individual bacterial cells on the agar surface.

To identify the organism: biochemical tests to identify bacteria are only valid when performed on pure cultures.

To produce isolated colonies of an organism (primarily bacteria) on an agar plate. This is useful when we separate organisms in a mixed culture (to purify/isolate a particular strain from contaminants) or to study an organism’s colony morphology.

The purpose of the streak plate is to obtain isolated colonies from an inoculum. Isolated colonies represent a clone of cells derived from a single precursor.

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Make sure the surface of the plate is free of droplets of condensed moisture.

Flame the loop after you streak each quadrant.

Streak lightly so that you do not gouge the agar.

Agar plate ( nutrient agar or any other agar medium).

A source of bacteria (stock culture, previously streaked agar plate, or any other inoculum).

For example, clusters of staphylococci, chains of streptococci, etc. With species in which the cells form a characteristic grouping during cell divisions, the colony-forming unit may develop from a group of cells rather than form a single cell. Pure cultures can be obtained by picking well-isolated colonies and re-streaking these on fresh agar plates.Ī common assumption is an isolated colony of bacteria is the progeny of a single bacterial cell (i.e. An isolated colony is formed when these lone bacterial cells divide and give rise to thousands and thousands of new bacterial cells. While streaking in successive areas of the plate, the inoculum is diluted to the point where only one bacterial cell is deposited every few millimeters on the surface of the agar plate. The inoculum is diluted by streaking it across the surface of the agar plate.

Limitation of Streak Plate Technique Principle of Streaking.

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