MICROPARA Lab Exercise 2 DISINFECTION AND STERILIZATION

WESLEYAN UNIVERSITY-PHILIPPINES
Cushman Campus
Mabini Extension, Cabanatuan City Philippines, 3100
COLLEGE OF ARTS AND SCIENCES

MICROBIOLOGY AND PARASITOLOGY
NAME: _______________________________DATE: __________________________________
Exercise No. 2
DISINFECTION AND STERILIZATION
Objectives:
1. To acquire the different aseptic techniques being observed in the laboratory,
2. To learn to operate some laboratory equipment use in sterilization and disinfection.
Materials:
Test tubes, Petri dishes, hot air oven, inoculating needle/loop, sterilizer, autoclave, alcohol
lamp, chlorox (phenolic solution), 70% isopropyl alcohol, prepared agar media, sample of
pure culture.
Procedures
A. Aseptic Technique. Perform and draw the different aseptic technique describe below.
1. Surface Disinfection. Wash hands with antibacterial soap. Wet piece of cotton/ tissue
with isopropyl alcohol or Clorox and wipe the surface of your working area. Dispose
properly all the materials you are going to use.
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2. Flaming of inoculating loop/ needle. Pass the inoculating loop/needle over the hottest
portion of the non-luminous flame until the loop/needle becomes red hot. Hold this in
almost vertical position for maximum heating. Pass this for a few times over the flame.
Never put down or allow the loop/ needle to come into contact with any unsterilized
surface after flaming. Flaming should be done before and after using the loop/ needle
to kill the possible microorganism that adhere to it.
Allow the loop/needle to cool first using it in transferring or isolating microbial
culture.
3. Transferring inoculum from tube to tube (Petri dish)
A. Hold carefully the test tube containing the culture with one hand. Use the middle
and ring fingers of your free hand to remove the cotton plug. NEVER LAY THE
COTTON PLUG DOWN.
B. Hold the test tube in inclined position and raise the mouth of the tube about 1 to 2
inches above the flame. Pass it over the flame for a few seconds.
C. Insert a cooled sterilized inoculating loop/needle (performed from no. 2) inside the
tube to obtain a small amount of culture by touching gently the top of the media.
D. Draw the loop/ needle carefully. Be careful not to touch the sides of the tube while
doing this.
E. Flame again the mouth of the tube for few seconds before replacing the cotton plug
to the tube.

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F. Introduce the loop/ needle gently (do not touch the side of the tube) containing the
inoculum into another tube (Petri dish) by streaking the top of the media in a
zigzag pattern.
G. Cover the tube with cotton plug or the Petri dish with its cover.
H. Flame again the loop/ needle after using to kill the microorganism that adhere to it.
4. Inoculating the plate. Hold the Petri dish with one hand supporting the bottom with
your fingers. Using the little finger, try to rotate the plate allowing the side to pass over
the flame. Repeat this using the cover of the dish.


Cushman Campus

B. Sterilization
Familiarize yourself with one of the most commonly equipment to sterilize apparatus
in the laboratory. Draw and learn how to operate it.
The Hot Air Oven is the one available in our laboratory. Place dried glassware’s inside the
oven. Use temperature of 200 degree Celsius for fast heating. The sterilization process with
this temperature will be completed after 3 hours. Reserve the sterilized
Building people,glassware’s
Transforming for next
lives.
laboratory period. Some sterilizer are used to sterilize
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laboratory gown,
Admin@wesleyan.edu.ph cotton, tissue,
(044)463-2162/4632074 gauze,
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needle, syringe, etc. such as those used in hospitals.

Cushman Campus

Discussion:
1. State the differences between disinfection and sterilization.
2. Give instances that disinfection and sterilization are applied in your everyday
life.

3. Why is it important that the inoculating loop/ needle must be cooled first before
allowing coming into contact with the culture?

Cushman Campus



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WESLEYAN UNIVERSITY-PHILIPPINES

Mabini Extension, Cabanatuan City Philippines, 3100

COLLEGE OF ARTS AND SCIENCES

Building people, Transforming lives.

GEN-FM-019 Rev 1 Effective 30 Sept 2022

Mabini Extension, Cabanatuan City Philippines, 3100

COLLEGE OF ARTS AND SCIENCES

3. Transferring inoculum from tube to tube (Petri dish)

GEN-FM-019 Rev 1 Effective 30 Sept 2022

Mabini Extension, Cabanatuan City Philippines, 3100

COLLEGE OF ARTS AND SCIENCES

Building people, Transforming lives.

GEN-FM-019 Rev 1 Effective 30 Sept 2022

Mabini Extension, Cabanatuan City Philippines, 3100

COLLEGE OF ARTS AND SCIENCES

GEN-FM-019 Rev 1 Effective 30 Sept 2022

Mabini Extension, Cabanatuan City Philippines, 3100

COLLEGE OF ARTS AND SCIENCES

Building people, Transforming lives.

GEN-FM-019 Rev 1 Effective 30 Sept 2022

Mabini Extension, Cabanatuan City Philippines, 3100

COLLEGE OF ARTS AND SCIENCES

Building people, Transforming lives.

GEN-FM-019 Rev 1 Effective 30 Sept 2022

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