Why single hyphal tipmethod is used than spore suspension?
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1. Prepare a dilute spore suspension in 1% aqueous glycerine or in water to which a small amount of detergent has been added.
2. Using a sterile loop, remove a loopful of the spore suspension and place it on a slide. Examine the slide to determine relative concentration and appearance of the spores.
3. Using a sterile loop, streak another loopful of the spore suspension over the surface of agar in Petri dish (both agar and Petri dish should be sterile). Sterilize the loop and re‐streak (without additional inoculum) in the opposite direction.
4. Place the Petri dish on stage of the microscope and, using low power, locate the spores, or allow the sample to germinate overnight, and then locate them.
5. After single spores have been located, use a sharp sterile needle to cut out a square of agar containing the spore and transfer it (agar and spore) to a fresh sterile agar plate or test tube slant.
2. Using a sterile loop, remove a loopful of the spore suspension and place it on a slide. Examine the slide to determine relative concentration and appearance of the spores.
3. Using a sterile loop, streak another loopful of the spore suspension over the surface of agar in Petri dish (both agar and Petri dish should be sterile). Sterilize the loop and re‐streak (without additional inoculum) in the opposite direction.
4. Place the Petri dish on stage of the microscope and, using low power, locate the spores, or allow the sample to germinate overnight, and then locate them.
5. After single spores have been located, use a sharp sterile needle to cut out a square of agar containing the spore and transfer it (agar and spore) to a fresh sterile agar plate or test tube slant.
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