Lab Report 2

Bacterial Culture Techniques

- BIO 223 L -

Student Name: Sara Alabdulkarim

Student ID: 201136
Submission date: September 26, 2021

Abstract:

Objective:

To transfer bacteria between different types of media cultures in an aseptic manner

and to learn how to isolate a single species in pure culture by plate streaking
technique.

Introduction:

Materials:
 Nutrient broth and nutrient agar slant culture.
 Nutrient agar plates.
 Agar slant, broth tubes.
 Disposable inoculating loops.
 Flame.
Procedure:
(A) Aseptic Transfer Techniques:

I. From Broth to Agar slant:

1. Flame the mouth of the broth tube before you start.

2. Insert the loop into the broth culture and withdraw a loopful of liquid. You
should see watery film on the loop and re-flame the mouth of the tube.
3. The broth-containing loop is immediately inserted into the end of the agar
slant tube.
4. Place the loop flat on the agar surface and move the loop rapidly in a zigzag
motion, gently across the agar surface from bottom to top.
5. Remove and dispose the loop.

II. From Agar plate to Broth:

1. Flame the mouth of the broth tube before you start.

2. Insert the loop into the agar plate culture and touch the agar surface where
the bacteria are growing.
3. Remove a minute sample of bacteria.
4. Insert the loop is immediately into the broth tube.
5. Shack the loop gently to suspend the bacteria.
6. Withdraw the loop from the broth and dispose it.

(B) Quadrant Streak plate Technique:

1. Select an agar plate and label the bottom side of the plate by group number
and the date.
2. Obtain a broth culture and Flame the mouth of the broth tube before you
start.
3. Insert the loop into the broth culture and withdraw a loopful of liquid and re-
flame the mouth of the tube.
4. Streak the loop (zigzag motion) along one area/quadrant of the plate.
5. Pick a new loop and streak some of the bacteria from the first area to the
second area.
6. Repeat step 5 on the third and fourth areas. Replace the lid.
Invert the plate and incubate it for 24 h at the appropriate temperature
 Results:

Figure 1: Agar plate with growing bacteria . Figure 2: Agar slant with bacterial cultures.

Figure 3: Agar broth, slight bacterial growth.

Discussion:
In this lab we preformed Aseptic Transfer of bacteria between different types of
media culture. First one as an agar plate where we took a loopful of specimen and
transferred it on to the surface or the plate by streaking a series of parallel lines in
different parts of the plate that resulted in separated colonies over the streaked
segments. Second one was the broth where we picked up a segment of the bacteria
and twirled it around in the broth and results were that the broth got “foggy”. As for
the third and last one (agar slant) we again to a segment of the bacteria and dragged
the loop in a ridged line across the slant, result where that the bacteria appeared on
the surface across the slant ridged line we drew.

References:
o Lab manual.
o Lab PowerPoint.
o