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Microbial Inoculation Procedure on Agar Slant Medium

Agar slant medium is a commonly used culture medium in microbiology for the isolation and preservation of bacterial cultures. This method allows for easy observation of colony growth and maintains the viability of microorganisms over time.

(1) Materials

• Agar slant medium
• Cultured plate with isolated colonies
• Inoculating loop or needle
• Flame source (Bunsen burner)
• Cotton plugs
• Tube rack

(2) Method

1. Label the upper part of the agar slant test tube with the name or number of the transplanted seed, the date of inoculation, the inoculator's group, and the code. This helps in tracking and identification during future experiments.
2. Hold the bacterial plate in your left hand and the inoculating loop in your right. Sterilize the loop by passing it through a flame until it glows red, then allow it to cool slightly. Carefully pick up well-developed isolated colonies from the plate using the loop. Place the plate back in its original position.
3. Hold the agar slant tube diagonally with your right hand. Use your little finger and palm to hold the cotton plug. Gently rotate and remove the plug, then sterilize the tube mouth by passing it through the flame. Insert the inoculated loop into the agar at the bottom of the tube, and draw a straight line along the surface of the slant. Continue with a zigzag pattern from the base, or form a curved line if desired. Remove the loop, sterilize the tube again, replace the cotton plug, and place the tube in the rack. Finally, sterilize the inoculating loop by flame and put it down.
4. If the bacteria were grown in a liquid medium, hold the slant tube and the strain tube together with your left hand. Use your right ring finger and little finger to hold both cotton plugs. After sterilizing the tube mouth and the loop, take the bacteria from the strain tube and immediately transfer them to the slant surface. Replace the cotton plug and sterilize the loop once more before placing it back. Incubate the slant medium in a 37°C incubator for 24 hours. After incubation, a uniform lawn of bacterial growth should appear on the surface of the slant.

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