Pure Culture

Technique

The term "Pure Culture" refers to a population

or colony of cells growing in the absence of
other species or types. The idea that is if an
agar plate is inoculated with only one species
and there is no contamination, then each colony
on a plate will consist of genetically identical
cells have have come from a single progenitor
or parent cell. 

​Pure culture technique allows us to isolate one

Pure
species  from a mixed culture is a useful tool
that helps to obtain a single kind of organism
from a mixed culture. 

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There are two common methods of pure
culture. Both of these methods result in
individual colonies that are isolated
from a mixed culture.
culture. 

1. Pour Plate - pour plate

2. Streak Plate. 

H ow to Distinginguish 

1. The color of the colony

Empuls by Xoxoday
Truely connect with your team
2. Gram stain results. 

​  Mixed culture of the following

•
organisms
o Staphylococcus o Chromobacteriu o​  Serratia

aureus 
m violaceum
marcescens 

​ Gram-po sitive Gram-negative ​Gram-negative

(+) Cocci (-)
(-)

(spherical )
​ Bacilli (rod- Bacilli (rod-
ream /yellow shaped )
shaped )

colonies  blue /purple RED colonies

colonies

S. aureus bacterial colonies


STREAK PLATE
METHOD


 By Brooke Bearden - Own work, CC BY-SA 4.0,
https://commons.wikimedia.org/w/index.php?curid=64501189


 By Bill Branson – (Edited by Fir0002)(Edited by Drhx) -
This image was released by the National Cancer
Institute, an agency part of the National Institutes of
Health, with the ID 2230 (image) (next).
By Bill Branson
– (Edited by Fir0002)(Edited by Drhx) - This image was
released by the National Cancer Institute, an agency part
of the National Institutes of Health, with the ID 2230
(image) (next). Public Domain,
https://commons.wikimedia.org/w/index.php?
curid=3346760

Pour Plate Method

The pour plate technique

1 
 3

*You only have one shot at getting this

Mixed
right, because the TSA agar will solidify
Culture in quickly. Have everything completely
Broth
prepared and "ready to go" before
obtaining your TSA tubes.

1. Obtain 3 empty plates (Petri

dishes).
2. Label them 1, 2, and 3 along with, ​
names and lab days and time and
organism.
3. Obtain the broth containing the
mixed bacterial culture and a
loop, and flame the loop.
4. Retrieve one melted TSA tube
(TSA tube 1) from the water bath
and immediately transfer one
loopful of the mixed culture broth
to the tube of melted TSA (TSA
tube # 1)
5. Use loop to mix 5 seconds.
6. Have your partner
retrieve another melted TSA tube
(TSA tube 2) from the water bath
and immediately  transfer one
loopful of the mixture from the


first melted TSA tube  (TSA tube 1)
 to the second melted TSA
tube (TSA tube 2).
7. Use the loop to mix 5 seconds.
8. Immediately pour the contents of
TSA tube 1 into the petri plate
labeled 1.
9. Have your partner obtain a 3rd
melted TSA tube from the water
bath (TSA tube 3)
and immediately  transfer one
loopful of the mixture from the
second melted TSA tube  (TSA tube
2) to the third melted TSA
tube (TSA tube 3).
10. Immediately pour the contents of
TSA tube 2 into the petri plate
labeled 2.
11. Immediately pour the contents of
TSA tube 3 into the petri plate
labeled 3.  
12. Gently rotate the plate to help the
media spread out before
solidifying. 
13. After your plates have solidified,
incubate them upside down to
avoid condensation.

Why are Petri dishes

stored upside-down ?

   Petri plates are incubated upside-

down to lessen the risk of
contamination from airborne
particles settling on them and to
prevent the accumulation of any
water condensation that may

otherwise disturb or compromise a
culture .

After Incubation

• You will need your streak plates and pour plates and three TSA
slants for isolating your colonies. 

Ideally, you should see distinct separate colonies that are RED,
PURPLE and WHITE.
Isolate one of each colored colonies to its own dedicated slant tube
and incubate at 25 degrees C.  

After incubation, do a smear prep of the contents of each of your

slants and perform a Gram-stain. 

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