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Exercise 1-4 Micro Lab: Streak Plate Methods of Isolation

front 1

mixed culture

back 1

a microbial culture consisting of two or more species

front 2

pure culture

back 2

a microbial culture consisting of only a single species

front 3

What is the first step in identifying an organism?

back 3

isolation technique

front 4

isolation technique

back 4

obtaining isolation of individual species from a mixed sample

front 5

A commonly used isolation technique

back 5

streak plate

front 6

streak plate method of isolation

back 6

a bacterial sample (a mixed culture) is streaked over the surface of a plated agar medium. the cell density decreases, eventually leading to individual cells being deposited separately on the agar surface.

front 7

colonies

back 7

cells (on the streak plate that have been sufficiently isolated) that consist of only the original cell type grow into colonies

front 8

colony forming unit (CFU)

back 8

some colonies originate from individual cells and others from pairs, chains, or clusters of cells

front 9

What are streak patterns designed to do?

back 9

to separate deposited cells (CFUs) on the agar surface so individual cells (CFUs) grow into isolated colonies.

front 10

quadrant streak method

back 10

usually perfomed as the initial streak for isolation of two or more bacterial species in a mixed culture with samples suspected of high cell density

front 11

zigzag streak pattern

back 11

usually used with samples containing lower cell densities and with pure cultures when isolation is not necessary

front 12

The identification process of an unknown microbe relies on what?

back 12

obtaining a pure culture of that organism

front 13

How do you create a pure culture?

back 13

a portion of an isolated colony is transferred to a sterile medium to start a pure culture

front 14

When performing streak plate inoculation, what are the two ways you can hold the plate?

back 14

1. leave the sterile agar plate on the table and lift the lid slightly, using it as a shield from airborne contamination

2. place the plate lid down on the table and then remove the base and hold it in the air on an angle

front 15

What are the 4 steps in the quadrant streak method using a loop?

back 15

1. streak the mixed culture back and forth in one quadrant of the agar plate. flame the loop.

2. rotate the plate 90 degrees and touch the agar in an uninoculated region to cool the loop. flame the loop.

3. rotate the plate nearly 90 degrees and streak again using the same wrist motion. cool loop prior to streaking. flame loop.

4. after cooling the loop, streak one last time into the center of the plate. flame the loop and incubate the plate in an inverted position.

front 16

What 2 broth cultures will we use in class for practicing streaking methods?

back 16

1. micrococcus luteus
2. staphylococcus epidermidis

front 17

What 2 types of agar plates are generally used?

back 17

1. Trypticase soy agar
2. Nutrient agar plate