Activity 4 - Streaking GR 4

Office of the College of Arts and Sciences
Group No. 4 Section: BS BIO - 4203

Members:
Robino, Champagne B.
Sangalang, Jonah Mae N.
Virtucio, Nickole Andriea M.
Yu, Dolleen Quisha B.
INTRODUCTION
Streak plating is a fundamental technique used in microbiology to isolate and culture

bacterial colonies from a mixed population. There are various types of streak plating methods,
including quadrant streaking, T-streaking, and zigzag streaking, each tailored to specific
experimental needs.
Streak plating is crucial in microbiology for several reasons. First, for isolation of pure
cultures, by streaking, individual bacterial cells are separated, allowing for the growth of pure
colonies that can be studied and characterized. Second, for identification of microorganisms.
Pure colonies obtained through streak plating can be further analyzed using various biochemical
and molecular techniques for species identification and characterization. Last, for the study of
microbial diversity. Streak plating enables researchers to study the diversity of microorganisms
present in a sample by isolating and identifying different bacterial species.In conclusion, streak
plating is an essential technique in microbiology for isolating pure cultures and studying
microbial diversity, facilitating various research and diagnostic applications in the field.
OBJECTIVES
The objectives of this laboratory activity are the following:
● To practice how to effectively streak with avoidance of contamination

● To master streaking in a four-quadrant with ease
● To practice the four different isolation and inoculation techniques (plate to plate, plate to
tube, tube to tube, and tube to plate)
MATERIALS
● Petri Dish
● Test Tube
● Agar
● Inoculating loop
● Alcohol lamp
METHODS
The following is the procedure for our streaking activity, highlighting the important preparation
and tube-to-plate technique only. Stated below are the different isolation and inoculation techniques
(Isolation to point A to inoculation to point B) that can be used in many industries of microbiology:
1. Plate to Plate
2. Plate to Tube
3. Tube to Tube
4. Tube to Plate
Where…
• Slants (test tube)
- Best for preservation
- Oil makes the medium more long-lasting to be preserved3
- Easy to store
• Plates
- Initial inoculation
STREAKING PROCEDURE
(w/ reminders)
1. Prepare your growth medium
2. Practice Streaking • Start at the top of the plate

• Preferably when streaking, the
glass sound will be heard as it
Important
means you maximize the spaces
Preparation in
Streaking effectively, just make sure that
your pacing in streaking is just
right.
2.1 Old streaking: Using
muscle memory (faster and
traditional)
2.2 New streaking: 16-streak
(slower but high-end result and
more efficient)
3. Prepare all necessary materials and

equipment for streaking.
Tube to plate
4. Light the alcohol lamp and sterilize the Remember: The blue flame is
inoculating loop until it's glowing red hot. hotter than the red, so it's better
to use the blue part flame for an
inoculating loop
5. Let the inoculating loop cool for about 10-15 • If you doubt if it's hot or cold
seconds before transferring any microbes. you can test it, by tapping at the
side part of your culture media.
Regardless if it sizzles or not
don't put it back on the flame
since the sizzles already cool
down the inoculation loop.
6. Obtain microbes from the test tube (slant) 6.1 Use the ring and pinky
finger (top of knuckle) to
remove the cotton plug
6.2 Heat the mouth of the test
tube
6.3 Not too thick, since this
could affect the final result.
6.4 Flame the mouth
6.5 Bring back the cotton plug
7. Flame the mouth of the plate where you'll

transfer the obtained microbes
8. Streaking 8.1 Start at the top of the plate

8.2 Streak at the first quadrant
(initial streaking) (contains a
high concentration of inoculum)
8.3 Flame the mouth of the plate
8.4 Rotate the plate
counterclockwise (thumb at the
top of the plate, index finger
moving the plate, and the rest of
3 fingers holding the plate at the
bottom)
8.5 Sterilize the inoculating loop
until it's glowing red hot.
8.6 Then proceed to the second
quadrant where you'll obtain and
spread the inoculum from the
first streak in the first quadrant
8.7 Proceed to the third quadrant
and fourth quadrant performing
the same steps.
9. Store the plate or test tube (slant) at

appropriate storage with the right temperature
10. Don't forget to wash the laboratory

equipment and clean the respective used area.
DISCUSSION
The streaking method is used for obtaining pure culture. This technique is used for
isolating the bacteria and other microorganisms from a mixed population into a pure culture. This
process dilutes the bacteria, by streaking the inoculum over the agar surface to “thin out” the
bacteria, allowing individual colonies to grow separately. This method must be performed
properly because once the bacteria are spread too close together on a plate, they grow into a thick
layer, which is called confluent growth. This makes it hard to see individual colonies and fails to
grow a pure culture bacteria.
In this laboratory activity, the students performed different kinds of streaking methods
such as plate-to-plate streaking, plate-to-tube streaking, tube-to-tube streaking, and tube-to-plate
streaking. The students learned how to execute the aseptic technique and the proper way to hold
the agar plate, inoculating loop, and cotton plug. We also learned the different streaking patterns,
the old-fashioned way, and the new version, the 16s streaking method. As the provided time is
long, therefore, the students will be able to try numerous times to perform the different kinds of
streaking methods and the different streaking patterns. Therefore, the students became more
familiarized and built the skill for streaking.
CONCLUSION
The streaking method, which is essential in microbiology, allows the separation of pure
cultures from heterogeneous populations of microorganisms. This approach allows for the
development of individual colonies on agar plates by precisely manipulating the inoculum,
which is critical for subsequent analysis and characterization. In this laboratory activity, students
performed numerous streaking strategies to improve their aseptic abilities and familiarity with
laboratory equipment. Exposure to various streaking patterns improved their comprehension of
maximizing space on agar surfaces while maintaining optimum microbial dilution. Finally, this
exercise provided students with the required abilities for successful streaking, equipping them to
isolate and analyze microorganisms successfully in future research and diagnostic applications
that they may employ in their chosen career path.

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Office of the College of Arts and Sciences

Group No. 4 Section: BS BIO - 4203

Streak plating is a fundamental technique used in microbiology to isolate and culture

The objectives of this laboratory activity are the following:

● To practice how to effectively streak with avoidance of contamination

2. Practice Streaking • Start at the top of the plate

3. Prepare all necessary materials and

7. Flame the mouth of the plate where you'll

8. Streaking 8.1 Start at the top of the plate

9. Store the plate or test tube (slant) at

10. Don't forget to wash the laboratory

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