Purification and maintenance of microbes
Introduction
The process of purification and maintenance is to obtain only one type or
strain of microorganism from a mixed microbial population. In some bacteria,
we often need to purify a particular target bacterium because multiple
species are often present in the sample. Through purification, we are able to
evaluate the microbiological data of the specimen, which is often used for
contamination identification, active substance testing, product quality
parameter evaluation, etc.
Streak plate
AIM
To purify microorganisms from a mixed Population.
PRINCIPLE
It is also called Looping out method. It is not a proper quantitative method
for the isolation and enumeration of bacteria and other microorganisms. It is
a good method for microbial population. This method is called streak became
it creates lines on the surface of the medium like streaking . Inoculation loop
is used to transfer inoculum from the sample for streaking and is called
looping out technique.
Different types of streaking techniques are available, they are T-streak,
quadrant Streak, Simple streak and continuous streak. These techniques are
named based on the type of streak drawn on the surface of the medium.
MATERIALS REQUIRED
Sample of mixed culture
Nutrient agar
Petri plates
Autoclave
Inoculation Loop
PROCEDURE
1. Prepare nutrient agar or any required medium and pour in the plates.
2. Allow the plates to solidify.
3. Sterilize the inoculation loop using flaming technique.
4. Transfer microbial mixture from a tube to the edge of an agar plate
with an Inoculation loop.
5. Incubate the plates at 37°c for 24 hrs.
Pour plate
AIM
To isolate microorganisms from the sample
To get pure culture of microorganisms
PRINCIPLE
Pour plate is a rapid quantitative isolation method. This method is used to
isolate bacteria, fungus and actinomycetes. Original sample is Diluted
Several times to thin out the population sufficiently. Then most diluted is
mixed with warm agar and poured in the petri plates. After the agar is
hardened, each cell is fixed in the plane and forms an individual colony. The
total number of colonies equals the number of viable microorganisms. Co
MATERIALS REQUIRED
1. Nutrient agar
2. Test tubes
3. Petri plates
4. Autoclave
5. Pipettes
PROCEDURE
1. Prepare nutrient agar and sterilize at 121°C for 15 mins.
2. Dilute the sample upto 10-7 using diluent.
3. Add 1ml of Sample from 10- 3 dilution to approximately labelled petri
plate 10-3
4. Pour the medium in to sample added Petri plates
5. Rotate the Petri plates clockwise and anticlock wise.
6. Allow the plates to solidity.
7. Similarly perform pour plating for other dilutions like 10-4, 10-5, 10-6.
8. Incubate all nutrient agar plates at 37°C for 24 hours and record the
results.
