1. Taking care not to touch the cotton wool end, remove a sterile swab from the pack and hold it in your right hand.
  2. Lift a bottle of sterile water in the left hand. Do not put down swab.
  3. Remove the lid of the bottle with the little finger of the right hand which is still holding the swab.
  4. Flame the neck of the bottle.
  5. Insert and withdraw the cotton wool bud.
  6. Flame the neck of the bottle and replace the lid.
  7. Rub the swab on the area to be tested for 10 – 15 seconds.

image 2        image 1

  1. Loosen the top of the bottle containing the slope culture so that it can be removed easily.  loosen top of bottle
  2. Hold the loop in the right hand.
  3. Flame the loop and allow to cool. Do not put down loop or wave it around.
  4. Lift the universal containing the inoculum with the left hand.
  5. Remove the lid of the universal with the little finger of the right hand. (Turn the bottle, not the lid). Do not put down the lid.
  6. Flame the neck of the universal.
  7. remove a small inoculum Bringing the bottle to the loop, insert the loop into the bottle, touch a small area of growth and withdraw loop.  touch a small area of growth  
  8. At all times, hold the loop as still as possible.
  9. Flame the neck of the universal.
  10. Replace the lid on the universal using the little finger, turning the bottle not the lid.
  11. Place slope on bench.

image4

    1. Hold the loop in the right hand.
    2. Flame the loop and allow to cool. Do not put down loop.
    3. With the left hand, lift the lid a little of the Petri dish containing the inoculum.
    4. Touch a single colony with the wire loop. 
    5. Withdraw loop carefully without touching plate. Do not put down loop or wave it around.
    6. Replace lid of Petri dish.
  1. Loosen the tops of the universals containing the broth cultures so that they can be removed easily.
  2. Remove the pipette from its sterile container taking care not to touch the tip, attach bulb and hold in the right hand.   
  3. Lift the universal containing the inoculum with the left hand.
  4. Remove the lid of the universal with the little finger of the right hand. (Turn the bottle, not the lid). Do not put down the lid. 
  5. Flame the neck of the universal.
  6. Squeeze the teat of the pipette before it enters the broth so that it does not cause bubbles and possible aerosols and withdraw a little of the culture.   
  7. At all times, hold the pipette as still as possible.
  8. Flame the neck of the universal.
  9. Replace the lid on the universal using the little finger, turning the bottle not the lid.
  10. Place universal on bench.

image 5

Take care that the culture does not drip from the pipette. Work over paper towels moistened with disinfectant if working with novices.

image 6

  1. Dip the implement in ethanol, place in Bunsen flame briefly and allow ethanol to burn off. Do not put down.
  2. Lift the lid a little of the Petri dish containing the inoculum with the left hand.
  3. Hold the inoculating implement in the right hand.
  4. Keep the ethanol at a safe distance from the Bunsen flame.
    1. with scalpel, cut a square shape in the fungal mycelium;
    2. with forceps, pick up a little piece of mycelium;
    3. with cork borer or pipette, cut a core of mycelium;(if the centre of the colony is producing spores, remove the inoculum from the edge of the colony).  mould colony showing circular 'stamps' ready for extraction
  5. Lift a cube, strands or core of agar with fungal mycelium.  picking off a circular inoculum
  6. Withdraw implement from Petri dish.
  7. Replace lid of Petri dish.
  8. At all times, hold the inoculating implement as still as possible.