Bret Lundgren English 202C

How to Grow Bacteria in a Petri Dish

Anyone in a profession that incorporates knowledge from biology will tell you that knowing
how to grow bacteria in a petri dish is one of the most fundamental skills one gains from a
biology class. This information can be used in any job or research position that involves
biological study of bacteria. Growing bacteria in a petri dish is easy to do with versatile
applications in fields such as microbiology and immunology. Although the protocol is fairly
simple, first learning how to grow bacteria can be confusing without some guidance. This
instruction set will be useful in helping you, a high school Biology II student, learn how to grow
bacteria in a petri dish. The overall process should take approximately 3 to 5 days to complete.

WARNING: GROWING BACTERIA IN A PETRI DISH CAN

BE DANGEROUS IF INFECTIOUS/HARMFUL BACTERIA
ARE CULTURED AND NOT STORED AND DISPOSED OF
PROPERLY. USE PROPER LAB TECHNIQUES WITH
TEACHER SUPERVISION.

Background:
Bacteria are unicellular (single-celled) microorganisms. They are so small that they cannot be
seen by the naked eye, but they reproduce quickly, forming a colony that can be seen by the
naked eye. Bacteria reproduce and form colonies through binary fission – the process by which
a bacteria cell splits into two cells. This rapid growth is a useful feature when using bacteria as a
model organism for an experiment or lab analysis. Bacterial growth in a lab occurs in a petri
dish (a shallow, circular, transparent dish) on an agar gel, as shown in Figure 1. Agar gel is a
jelly-like substance that is used to culture bacteria, as it has an abundance of nutrients that are
required for bacterial growth.
*The whole procedure takes about 3 to 5 days to complete.

Figure 1: Bacteria Grown in Petri Dishes

Bret Lundgren English 202C

Materials:
 2 Petri Dishes with Lids  Heat Resistant Gloves
 500 mL Beaker  Lab Apron
 ½ Teaspoon Agar  Goggles
 ¼ Cup Hot Water  Bleach
 Hot Plate  Masking Tape
 Magnetic Stirrer  Ziploc Bag
 2 Sterile Cotton Swabs  Bacterial Source

*These materials and instructions are based on a process that allows for two petri dishes to have
bacterial cultures. Adjust the proportions accordingly in order to make more or less cultures.

Procedure:
**Wear lab apron and goggles throughout each step in the experiment. Wear heat resistant
gloves when dealing with the hot plate and any hot liquids/materials.

Step 1: Making the Agar

*Average Time for Step 1: 20 Minutes
1. Obtain and boil ¼ cup of water in the 500 mL beaker on a hot plate, as shown in Figure 2

Figure 2: Water Boiling in

500 mL Beaker on Hot Plate

Figure 3: Boiled Water Being

Stirred by Magnetic Stirrer

2. Put the magnetic stir bar in the beaker on the hot plate and turn it on
2.1 If the magnetic stirrer is functioning properly, it should resemble Figure 3
3. Slowly and gradually add the ½ teaspoon of agar to the boiling water
4. Let the magnetic stirrer mix until there is a consistent mixture with no particles or
chunks floating
4.1 DO NOT LET THE AGAR SOLUTION BOIL OVER
Bret Lundgren English 202C

5. Remove beaker with agar solution from hot plate

6. Let agar mixture cool for about 3 to 5 minutes
6.1 Do not let agar mixture cool too long or it will solidify in the beaker
7. Pour the cooled agar mixture into the petri dishes so that each one is half filled, as
shown in Figure 4

Figure 4: Petri Dish Filled Halfway

with the Cooled Agar Solution

8. Put lids on petri dishes

9. Let petri dishes cool until the agar solution solidifies into a gel

Step 2: Swabbing the Bacteria onto the Agar Gel

*Average Time for Step 2: 2 Minutes per Petri Dish
1. Take a sterile cotton swab out of the package and swab your bacterial source using all
sides of the cotton swab head
1.1 Note that the bacterial source is something that is chosen to test for the
presence of bacteria
2. Take the cotton swab and lightly swab the agar gel in the petri dish with the cotton
swab head (a useful image showing this pictorially can be seen in Figure 5)
2.1 Swab one quarter at a time, starting at the outside and working into the inside in
a zig-zag motion left-to-right
2.2 Swab from the middle back to the outside in the next quarter while in a zig-zag
motion.
2.3 Repeat this until you have swabbed all four quarters of the gel
2.3.1 These steps are all done while twisting the cotton swab head. Do
not lift the cotton swab head off of the agar gel during this entire
process
Bret Lundgren English 202C

Figure 5: Pathway of Swabbing

Agar Gel in Petri Dish

3. Put the lid back onto the petri dish and leave it sealed from this point forward

Step 3: Growing, Analyzing, and Disposing the Bacteria

*Average Time for Step 3: 3 to 5 days
1. Store the sealed petri dishes in a warm, dark place
2. Let the bacteria in the petri dishes grow for 3 to 5 days, until it resembles Figure 6
2.1 Check in on the bacteria once per day to observe growth progress and note any
changes
3. Once many colonies have formed and grown to a large size (about the size of the tip of a
pencil to a head of a cotton swab), take them out of their warm, dark place for analysis
3.1 Take detailed notes on the size, number,
appearance, color, and shape of the
colonies in each petri dish. Do not open the
petri dish at any point during the analysis
3.2 After your analysis is done, it will be time to
dispose of the petri dishes and colonies of
bacteria
4. Tape the petri dishes shut with masking tape so
they will not come open when discarded Figure 6: Petri Dish with
5. Place the petri dishes inside of a Ziploc bag Multiple Bacteria Cultures
6. Pour a large amount of bleach into the Ziploc bag
with the petri dishes
7. Seal the Ziploc bag and then discard properly according to the teacher’s instructions
7.1 This may include putting the Ziploc bag in a special disposal bin
8. Congratulations! You have successfully grown, analyzed, and discarded bacteria in a lab!
Bret Lundgren English 202C

Figure Sources:
1. https://www.stevespanglerscience.com/lab/experiments/growing-bacteria/
2. https://isenseproject.org/projects/1534
3. http://www.agarscientific.com/magnetic-stirrer-hotplates.html
4. https://commons.wikimedia.org/wiki/File:YPED_agar_plate.jpg
5. Sourced from Author
6. Sourced from Author