Streaking Method for Isolating Microorganisms
1. INTRODUCTION
Streaking is a popular scientific method for separating bacteria from a mixed
culture. This technique uses an inoculating loop to apply a tiny bit of the
culture to the surface of an agar plate. In order to help separate the germs
and allow for the growth of various colonies, the sample is diluted as you
streak. Because each colony starts from a single cell, studying certain
bacterial species is made simpler. For microbiologists, streaking is crucial
because it makes it possible to identify and characterize various organisms in
a sample, which is crucial in disciplines like environmental research, food
safety, and medicine.
2. OBJECTIVE
- Obtain pure cultures for further characterization and analysis.
3. MATERIALS / APPARATUS AND CHEMICALS
- Wire inoculating loop
- Glass slides
- Test tube rack
- Marking pen or pencils
- Beaker
- Erlenmeyer flask
- Petri dishes
- Laminar air flow hood
- pH meter
- Incubator
- Balance
- Autoclave
- Test tubes and holders
- Spatula
- Distilled water
- Refrigerator
- Bunsen burner
- Sterile swabs
- Alcohol for cleaning
- Nutrient agar
- Nutrient broth
- Selective media
- Dilution blanks (9 ml water)
- Blood agar plates
A mixed broth culture containing **Serratia marcescens** (pigmented),
**Escherichia coli**, and **Staphylococcus epidermidis** was used. A
demonstration plate culture made from this broth shows colonies isolated
using good streaking techniques.
4. PROCEDURE
1. Mix the content of the culture tube well.
2. Use the loop to pick up a loopful of culture broth and place it on the edge
of an agar dish without touching the edge. Lightly scratch the agar with the
loop.
3. Sterilize the loop and let it cool.
4. Rotate the agar dish in and draw four lines back and forth, each passing
through the inoculum and extending to one side of the agar plate.
5. Sterilize the loop again and let it cool.
6. Rotate the dish and draw another set of four lines, crossing the previous
ones and extending to the adjacent side.
7. Turn the plate over and repeat this parallel striping once more.
8. Finally, streak a few lines in the center of the plate without touching the
original inoculum.
9. Incubate the plate (inverted) at 35-37 °C.
10. After incubation, check the results for further analysis.
5. RESULTS
Fig.1.1 streaking from the petri dish At a concentration
−1
of 10
After incubating the agar dish at 35-37 °C, we observed the growth patterns
of the colonies. The streaking technique should result in isolated colonies,
with fewer cells seen as you move away from the original inoculation spot.
By looking at individual colonies, we can gather information about the
organisms present.
The degree of isolation achieved shows how well the streaking process
worked. Ideally, well-isolated colonies will be visible, indicating successful
dilution from the original sample.
CONCLUSION
The streak plating technique is a useful way to isolate pure cultures from
mixed populations of microorganisms. It helps separate individual cells that
grow into distinct colonies. Observing the characteristics of these colonies
can provide clues about the types of microbes present. However, further
testing—like microscopic examination, biochemical tests, or molecular
techniques—might be necessary for a definitive identification. Following
proper techniques throughout the streaking process, such as sterilizing loops
and avoiding contact with the original inoculum after the first streak, is
crucial for reliable results. Isolated colonies should be chosen for further
testing (like Gram staining or metabolic tests) to gain deeper insights into
the microorganisms and their potential uses in research, industry, or
healthcare.
