STANDART OPERATING PROCEDURE FOR BACTERIAL BREEDING WITH

SPREAD PLATE METHODON LABORATORY OF THE HEALTH ANALYSIS

DEPARTMENT OF HEALTH POLYTECHNIC OF SEMARANG

Number : SOP/0335/POLKESMARLAB
Date : 23 April 2020
Provided by : Health Analysis Department of Health Polytechnic of Semarang

1. Purpose
To increase the number of bacteria by breeding them so that they can be used in
practical activities in the laboratory of the Health Analysis Department of Health
Polytechnic of Semarang
2. Scope
Laboratory users who will breed bacteria. including students, lecturers, laboratory
assistants, staff, and employees of the health analyst department of the Semarang
Health Polytechnic
3. Responsibility
Anyone who enters the laboratory of the health analyst department is responsible for
implementing procedures on how to breed bacteria properly
4. Definition
a) Bacteria are microscopic organisms that are not visible to the naked eye. They
can live in diverse environments, both inside and outside the human body
(such as land, river water, and sea water).
b) Bacterial breeding is a method used to produce new bacteria which will later
be able to be used for practicum activities in the laboratory.
c) spread plate technique is a technique in growing microorganisms in the media
so that by pouring the bacterial culture stock or removing it on the media so
that it has solidified, while the culture pour plate is mixed when the media is
still liquid (not yet solidified).
5. Tools and Materials
a) Tools : stir bar, L rod, beaker glass, ose needle, petri dish, erlenmeyer,
measuring cup, incubator, bunsen, refrigerator, gauze, cotton, dropper,
micropipette, digital scale.
 b) Materials : aquadest, 70% ethanol, Nutrient agar (NA), bacterial culture.
6. Procedure
Making Nutrient Agar (NA) media
a) Weigh 7 grams of nutrient agar
b) Dissolve it into 250 ml aquadest in the erlenmeyer
c) Heat it on the stove while stirring until the solution becomes clear
d) Chill, then sterilize with an autoclave at 121oC for 15 minutes
e) Media that has been sterilized, can be cooled at room temperature
f) Keep in the refrigerator

Bacterial propagation by the spread plate method

a) NA media is inserted into a sterile petri dish and allow it to solidify
b) Drop 0.1 ml of bacterial culture onto the surface for which it has aseptically
solidified
c) Burn the L stem which has been previously dipped in alcohol, then cool
d) Spread the bacteria using the L stem evenly
e) Incubated bacteria with an incubator at 35 ±2oc
f) Bacterial growth was observed in the media
7. Results
Breeding Media type Bacteria The existence
method Culture type of a colony
Spread plate Nutrient Agar Staphylococus Yes, spread
method (NA) aureus evenly on the
surface of the
NA media

8. Reference
https://www.slideshare.net/dian0911/laporan-sterilisasi-pembuatan-media-dan-teknik-
inokulasi
https://aguskrisnoblog.wordpress.com/2011/01/14/teknik-membuat-biakan-murni/
http://fitri-fykatili.blogspot.com/2016/10/laporan-praktikum-pembiakan-bakteri.html