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Schematic Diagram (Pre Lab)

The document describes different plating methods for isolating bacteria including streak, pour, and spread plating. It discusses obtaining bacterial cultures, streaking or spreading them on nutrient agar plates, incubating the plates, and examining the bacterial growth. Morphological and cultural characterization of selected bacterial isolates is also covered.

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MIKAEL PATRICK DEIPARINE
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41 views

Schematic Diagram (Pre Lab)

The document describes different plating methods for isolating bacteria including streak, pour, and spread plating. It discusses obtaining bacterial cultures, streaking or spreading them on nutrient agar plates, incubating the plates, and examining the bacterial growth. Morphological and cultural characterization of selected bacterial isolates is also covered.

Uploaded by

MIKAEL PATRICK DEIPARINE
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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A.

ISOLATION OF BACTERIA USING THE PLATING METHOD

1. Streak Plating Method

PRINCIPLE METHOD:

obtain a small amount of mixed culture of bacteria

streak over the surface of a pre- solidified Nutrient Agar plate

STREAKING METHOD:

streak one loopful of organisms back and forth over Area 1

flame the loop, cool for 5 seconds

rotate the dish in a 90 degrees angle, remain closed

streak Area 2 back and forth, hitting the original streak

↓
flame the loop, cool for 5 seconds

rotate the dish

streak Area 3 back and forth, hitting the last area

flame the loop, cool for 5 seconds

rotate the dish

streak Area 4, contracting Area 3

drag out the culture


AFTER STREAKING:

invert the plate

keep inside an autoclavable plastic

incubate at 30C for 48 hours

AFTER INCUBATION:

document the growth pattern

describe the observations

2. Pour Plating Method

liquify Nutrient Agar, maintain at 50C

obtain 1 mL aliquots each from tubes containing the following dilutions 10-4,10-5
and 10-6.

place the inoculum in separate empty sterile plates in


duplicates

pour NA in each plate

mix the contents by moving its contents counter clockwise


↓

incubate at 30C for 48 hours

AFTER INCUBATION:

document the growth pattern

describe the observations

3. SPREAD PLATING METHOD

PRINCIPLE METHOD:

obtain 0.1 mL aliquots of the following dilutions 10-4, 10-5 and 10-6.

place the inoculum in separate NA pre-solidified duplicate plates

STERILIZATION METHOD:

sterilize an L-shaped glass rod in 95% ethyl alcohol

↓
flame until the alcohol has burned off

cool the L-rod by touching the side of the agar plate

ACTER STERILIZATION:

spread the inoculum over the surface of the agar using the cooled L-rod.

move L-rod on the agar plate

allow agar to solidify

invert the plate

keep inside an autoclavable plastic

incubate at 30C for 48 hours

AFTER INCUBATION:

document the growth pattern

describe the observations


B. CULTURAL CHARACTERIZATION OF SELECTED ISOLATES

describe the morphology of selected colonies (A and B) on the NA plates

determine the agar strokes characteristic

C. PREPARATION OF SPECIMENS FOR WET MOUNT

obtain a small amount of growth from an isolated colony in the SPM

place in a clean slide


↓

add a drop of distilled water

mix the specimen and the liquid

place the coverslip on top of the specimen

observe cells under the microscope using OIO

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