COLEGIO SAN AGUSTIN-BACOLOD
MLS 109 LABORATORY ACTIVITY MANUAL
Medical Technology Program
LABORATORY ACTIVITY #8-A
INOCULATION OF PLATED culture media (Streak Plate Method)
The manner of inoculating bacteria on agar plates follows specific patterns. These patterns
allow isolation of bacteria or allowing them to grow so one would know their cultural characteristics.
Most bacteriological work requires pure cultures or clones of bacteria. The isolation method most
commonly used to get pure cultures is the streak plate method. A sterile inoculating loop is dipped into
a mixed culture that contains more than one type of microbe and is streaked in a pattern over the
surface of the nutrient medium. As the pattern is traced, bacteria are rubbed off the loop far enough
apart to grow into isolated colonies. These colonies can be picked up with an inoculating loop and
transferred to a test tube of nutrient medium to form a pure culture containing only one type of
bacterium.
Desired learning outcomes
After performing this activity, students should be able to:
1. perform a streak plate technique to isolate discrete colonies for subculture;
2. understand the purpose of the streak plate technique; and
3. identify isolated colonies and differentiate between various colony types based on size,
shape, color, and other characteristics.
Materials
Inoculating wire loop
Alcohol lamp
Broth culture
Agar plates
Procedures
1. Preparation:
Ensure your work area is clean and organized.
Label the bottom of the agar plate with relevant information (e.g., date, type of medium,
organism, your initials).
2. Sterilization:
Light the Bunsen burner or alcohol lamp to create a sterile field.
Sterilize the inoculating loop or needle by passing it through the flame until it glows red-hot.
Allow it to cool for a few seconds.
3. Inoculum Preparation:
If you are using a broth culture, gently mix the broth by swirling the tube.
If you are using an agar slant or colony from another plate, ensure you have a small, well-
COLEGIO SAN AGUSTIN-BACOLOD
MLS 109 LABORATORY ACTIVITY MANUAL
Medical Technology Program
isolated colony.
4. Transfer of Inoculum:
Using the cooled, sterilized inoculating loop or needle, pick up a small amount of the
culture.
If you are using a sterile swab, dip it into the culture or moisten it with sterile saline/broth,
then touch it to the culture.
5. Inoculation of Agar Plate:
Streak Plate Method:
o Lift the lid of the agar plate slightly to avoid contamination.
o Starting at the edge of the plate, streak the inoculating loop back and forth across the
surface of the agar in a small section of the plate.
o Flame the loop, let it cool, and rotate the plate slightly. Streak the loop through the first
section and into a new section.
o Repeat this process to create several streaks, which will help to isolate single colonies.
6. Incubation:
Invert the inoculated agar plates to prevent condensation from dripping onto the agar
surface.
Place the plates in an incubator set to the appropriate temperature for the microorganism
being studied (e.g., 37°C for many human pathogens).
Incubate for the recommended time period (usually 24-48 hours).
7. Observation and Analysis:
After incubation, observe the plates for microbial growth.
Note the appearance of colonies (size, shape, color, etc.) and any other relevant
observations.
8. Disposal:
Properly dispose of used cultures and materials following your laboratory's biosafety
protocols.
Notes:
Always work near the flame to maintain a sterile environment.
Do not leave the inoculating loop in the flame for too long to avoid weakening the metal.
Ensure all materials are sterile before use to avoid contamination.
Follow your laboratory’s safety guidelines, including wearing appropriate personal
protective equipment (PPE).
Guide Questions
1. What is the primary purpose of the streak plate method?
2. Why do you need to flame the loop and cool it between each streaking step?
3. How can you address issues such as no growth or unexpected contamination?
COLEGIO SAN AGUSTIN-BACOLOD
MLS 109 LABORATORY ACTIVITY MANUAL
Medical Technology Program
Preparation of Streak Plate
Inoculating the Streak Plate
COLEGIO SAN AGUSTIN-BACOLOD
MLS 109 LABORATORY ACTIVITY MANUAL
Medical Technology Program
Streak Plate Patterns
COLEGIO SAN AGUSTIN-BACOLOD
MLS 109 LABORATORY ACTIVITY MANUAL
Medical Technology Program
Laboratory Activity #8-A
INOCULATION OF PLATED culture media (Streak Plate Method)
ACTIVITY SHEET
NAME OF STUDENT: ______________________________________ YEAR & SECTION: ______________
DATE PERFORMED: _______________________________________DATE SUBMITTED: ______________
Observation/Results
Instructions:
1. Paste a picture of the streaking pattern (1 example only) used in your streak plate
technique.
2. Paste a picture of the appearance (after incubation) of the colonies of bacteria
inoculated using your pattern in #1.
COLEGIO SAN AGUSTIN-BACOLOD
MLS 109 LABORATORY ACTIVITY MANUAL
Medical Technology Program
Discussion
Conclusion
Reference/s
