Pure Culture

mohamad alfitroh harun

 Pure culture/Isolation
of bacteria
Bacterial isolation is the taking or removal of microbes from their environment
in nature and growing them as pure cultures in artificial mediums. Bacteria
transferred from place to place must use aseptic procedures. Aseptic means
free from sepsis is a condition contaminated due to other microorganisms.
Aseptic techniques are especially important when working in contact with
bacteria. Some of the tools used to carry out this procedure are bunsen and
laminarair flow. If not executed properly, there is a possibility of contamination
by other myoorganisms that will interfere with the expected results. Aseptic
techniques also protect laboratories from bacterial contamination (Singleton
and Sainsbury, 2006).
 Bacterial
isolation methods
There are several methods for inoculating bacteria according to the type of medium
for which they are intended. In the medium to be upright, a puncture method is
carried out using an ose needle. On the medium to be tilted, a scratch method is
carried out using an ose loop. In petridish medium, streak plate method, pour plate
or spread plate method can be used. After inoculation, an incubation process is
carried out, which is storing the medium in a tool or container with a certain
temperature and period certainly, thus creating an environment that provides
suitable conditions for bacterial growth (Harley and Presscot, 2002). The
methods for isolating pure cultures of microorganisms according to Pelczar and
Chan (1986) are: - scratch cup, - stocking cup, -pour cup.
 Principles and
Purpose of Insulation

Isolation of a pure strain can be carried out in principle in stages. The first level is free to do manually
by diluting it as far as possible, often to 10-4 or 10-6.The second level is by media that is selective for
certain microbes or certain microbes that may still be in the same group. The third level of the colony,
which seems to be possible, still needs to be re-diluted or re-isolated in order to be more convincing.
proof that the isolate strains obtained are really pure strains (Mulyono, 1992).The working principle of
bacterial isolation is quite simple, namely by inoculating a small number of bacteria in a certain
medium that can constrain bacterial life.
The purpose of culture transfer is to master the technique of transferring bacterial cultures from one
container to another aseptically, so that only the expected pure culture grows. This is especially
important in the early stages of microbial isolation work, especially those derived from culture stocks
(not from substrates).
 Samples needed for
isolation
Before isolating, sampling is first carried out. Various sampling according to the place:
1. Soil sample
If the desired microorganism is in the soil, then the way of taking it is adapted to the
purpose. For example, if what is desired is rhizosphere microorganisms, then samples are
taken from around the roots near the soil surface.
2. Water sample
The retrieval technique is simply to open the lid of the petri dish containing sterile media
for ±5 minutes.

According to Jutono (1980), there are several ways to isolate

microbes. For this isolation, several important things must be
considered, including:
1. The properties of the microbial species to be isolated
2. Place of life or origin of the microbes
3. Medium for its suitable growth
4. How to plant such microbes
5. How to incubate these microbes
6. How to test that the isolated microbes have been pure cultures
and are in accordance with what is intended
THANK YOU...