Growing Bacteria in Petri Dishes

Growing Microorganisms
in Petri Dishes/Test Tube

• Microbial culture media preparation
is the process of mixing nutrients,
agents for buffering and
maintaining the osmotic balance, as
well as selective inhibitors or
indicators to create an agar or broth
that supports the growth and the
differentiation of microorganisms.
Microbial culture media should provide
optimal growth conditions for all or specific
types of microorganisms.
The precise composition of a medium

depends on the species being cultivated and
the application goal.
The medium pH needs to be adjusted

depending on the microorganisms.
Microbial culture media is classified based

on several parameters, like chemical
constituents, physical nature, and function.
Types of
culture media
Synthetic (Defined) medium is a media
with defined chemical commonly used to
culture photoautotrophs, such as
cyanobacteria and photosynthetic protists.
Classification It is widely used in research, to study
microorganism metabolism.
by chemical
composition Complex media is a media containing non-
defined chemical components, such as
peptone, meat extract, and yeast extract,
that meet the nutritional requirements of
different microorganisms.
Selective and differential
media are media based
Classification on either complex or
defined media
by chemical supplemented with
composition growth-promoting or
growth-inhibiting
additives
Materials
Petri dishes, agar, and sterile swabs
or an inoculating needle.
Nutrient Agar
• Supports the growth of various types of bacteria

and fungi and contains many of the nutrients
necessary for the growth of bacteria.
• Peptone, beef extract and agar.
• Tryptic soy agar- works well for fastidious
bacteria
An inoculation loop, also known as a smear
Inoculating loop, inoculation rod or microstreaker, is a
simple tool used primarily by microbiologists
loop to take and transfer a small sample
(inoculum) of a microorganism culture, for
example for stripping on a culture plate.
ASEPTIC
TECHNIQUES
Aseptic
technique
A method that involves
target-specific practices
and procedures under
suitably controlled
conditions to reduce the
contamination from
microbes.
•Before inoculation with the desired
microorganisms, microbiological media and all
materials coming into contact with it must be
sterile.
•During any subsequent handling of the bacterial
cultures, unwanted or contaminant organisms
must be excluded employing aseptic techniques.
•Sterilisation implies the complete destruction of
all microorganisms including spores.
Aseptic Procedures in the Lab
• Wear your lab coat and gloves.
• Tie back long hair.
• Leave all food and drink in your backpack. Do not
chew gum in lab.
• The only thing on your lab bench should be the
equipment you are working with and your lab book.
Place your backpacks on the floor where you or
someone else will not trip over them.
•The only thing on your lab table should be the
equipment you are working with and your lab
book. Place your backpacks on the floor where
you or someone else will not trip over them.
•Clean up all spills immediately!
•Decontaminate your lab table with disinfectant
such as (Ethyl Alcohol) before and after lab.
•Wash your hands before leaving the lab.
TRANSFERRING
ORGANISMS
•Avoid producing aerosols that you can
breathe in/ droplets.
•Transfer organisms from one tube to
another/ slides to plates quickly and safely.
•The longer your organism is exposed to the
air, the more opportunities there are for it
to get contaminated.
Broth to Broth Transfer
Procedures
•Usually when you are working with a broth
culture, the organisms will have settled to
the bottom
• Hold the tube near the top and flick the bottom of the tube with your other hand.
Continue flicking until the organisms are re-suspended.
• Place both your tubes in your left hand with the lids pointing
up.
•Place both your tubes in your left hand with
the lids pointing up.
• Hold the tubes closer to the bottom so that your hand
will not be close to the flame when you sterilize the
mouth of the tubes
Grab the inoculating loop far back on
the handle as if you were going to
write with it.
•Place your loop in the Bunsen burner briefly for
2 minutes to sterilize it.
Do not leave your loop exposed to the
burner for more than 5 minutes
•Sterilize the mouth of the tubes by passing
them through the flame 2-3 times.
This kills organisms on the opening of the
tubes.
•Hold the tubes slightly tipped to minimize
microorganisms in the air from falling into the
open tubes.
-Insert the loop into the cultured tube
without touching the sides of the tube.
• Carefully withdraw the inoculating loop without touching the
sides of the tube and insert it into the new tube without
touching the sides of the tube.
• When you withdraw your inoculating loop from the second,
freshly inoculated tube, be sure to touch it against the inside
of the tube to knock off excess fluid.
• Flame the mouth of your tubes and replace the cover.
• Place your loop in the Bunsen burner for 2 minutes to
sterilize it.
• Never lay your loop or other inoculating equipment on the
counter without first sterilizing it.

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Growing Microorganisms

in Petri Dishes/Test Tube

The precise composition of a medium

The medium pH needs to be adjusted

Microbial culture media is classified based

• Supports the growth of various types of bacteria

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