Laboratory Notes
Date:
Group Members:
Professor:
Laboratory Exercise 2: Microbiological Streak Plate Technique
Objective: To perform the streak plate technique for isolating pure microbial colonies from a mixed
culture using aseptic methods.
Materials Needed:
Nutrient agar plates
Inoculation loops (metal or plastic)
Bunsen burner/incinerator
Sterile specimen swabs (if applicable)
Personal protective equipment (PPE: lab coat, gloves)
Disinfectant (70% ethanol)
Marker for labeling plates
Procedure:
A. Preparation and Laboratory Setup
1. Workspace Preparation:
o Disinfect the lab bench with 70% ethanol before and after use.
o Organize materials within easy reach to maintain aseptic conditions.
o Wear PPE and practice hand hygiene.
2. Pre-Inoculation Steps:
o Allow the agar plate to reach room temperature before use.
o Label the bottom of the plate with sample information (name, date, sample ID).
o Avoid unnecessary exposure of the agar surface to prevent contamination.
B. Streak Plate Inoculation
1. Choosing the Inoculation Tool
Metal Loop: Requires flaming until red-hot, then cooling before inoculation.
Plastic Loop: Pre-sterilized, single-use.
Sterile Swab: Alternative for initial streaking from liquid samples.
Rueda Street, Calbayog City
063 055 5339857
Samar, Philippines 6710
[email protected] www.nwssu.edu.ph
2. Sterilization of Metal Loop
Flame the loop in the Bunsen burner until it glows red-hot.
Allow it to cool in ambient air (do not blow on it or touch surfaces).
Test for coolness by touching the sterile agar surface away from inoculation areas.
3. Four Quadrant Streaking Method
1. Quadrant 1:
o Dip the sterile loop into the sample.
o Gently streak a small section of the plate in a zigzag motion.
o Avoid digging into the agar.
2. Quadrant 2:
o Rotate the plate 90 degrees.
o Flame and cool the loop.
o Drag the loop from Quadrant 1 into Quadrant 2 with 2-3 passes.
o Streak lightly to thin out bacterial load.
3. Quadrant 3:
o Rotate the plate 90 degrees.
o Flame and cool the loop.
o Streak from Quadrant 2 into Quadrant 3.
4. Quadrant 4:
o Rotate the plate 90 degrees again.
o Flame and cool the loop.
o Drag a few streaks from Quadrant 3 into Quadrant 4.
o End with a light zigzag to maximize dilution.
C. Incubation and Observation
1. Plate Placement:
o Place the streaked plate in an incubator, media side up (lid on the bottom) to prevent
condensation contamination.
o Incubate at the recommended temperature for 24-48 hours.
2. Colony Observation:
o Examine the plate for isolated colonies.
o Avoid opening the plate unnecessarily.
o Identify single, well-separated colony-forming units (CFUs).
D. Common Mistakes and Best Practices
Mistakes to Avoid:
Over-inoculating, leading to excessive colony overlap.
Failing to sterilize the loop between quadrants.
Rueda Street, Calbayog City
063 055 5339857
Samar, Philippines 6710
[email protected] www.nwssu.edu.ph
Digging into or damaging the agar surface.
Contaminating the plate by improper handling.
Best Practices:
Maintain aseptic technique throughout the procedure.
Work efficiently but carefully.
Label plates clearly and handle them properly.
Keep a well-organized and sterile workspace.
E. Post-Laboratory Questions
1. What is the purpose of the streak plate technique?
2. Why is it important to sterilize the loop between streaking quadrants?
3. How can you tell if your streak plate was successful?
Rueda Street, Calbayog City
063 055 5339857
Samar, Philippines 6710
[email protected] www.nwssu.edu.ph
4. What are the consequences of not allowing the loop to cool before streaking?
Four Quadrant Streaking Method
Rueda Street, Calbayog City
063 055 5339857
Samar, Philippines 6710
[email protected] www.nwssu.edu.ph
6. Conclusion:
Summarize the key findings and what you learned from the lab. Discuss whether the objectives
were met.
Rueda Street, Calbayog City
063 055 5339857
Samar, Philippines 6710
[email protected] www.nwssu.edu.ph
