image 1

  1. Partially lift the lid of the agar plate and gently rub the swab across the surface of the agar (see diagram). Take care not to break the agar surface.  rub swab on plate
  2. Replace lid of Petri dish.
  3. Sellotape lid to base of plate (two diametrically opposed pieces).
  4. Place swab in discard jar.  place swab in discard jar

image 3

  1. Lift a sterile agar slope with the left hand.
  2. Remove the lid of the bottle/tube with the little finger of the right hand which still holds the charged loop.
  3. Flame the neck of the bottle/tube.
  4. Streak the loop backwards and forwards across the surface of the agar slope and remove the loop.
  5. Flame the neck of the bottle/tube, replace the lid and place the bottle/tube in rack or on bench.
  6. Flame the loop and place on a heat resistant mat.

image 5

  1. Partially lift the lid of the Petri dish containing the solid medium.
  2. Holding the charged loop parallel with the surface of the agar, smear the inoculum backwards and forwards across a small area of the medium ( streaked area = A ).
  3. Flame the loop and allow to cool. Turn dish and streak loop from region A across the surface of the agar in three parallel lines ( to B ).
  4. Flame the loop and allow to cool. Turn dish and streak loop from B across the surface of the agar in three parallel lines ( to C ).
  5. Turn dish and streak loop from C to D across surface of agar.
  6. Replace lid of Petri dish.
  7. Flame loop and place on heat resistant mat.

image a               image b                  image c

image

  1. With the left hand, partially lift the lid of the Petri dish containing the solid medium.
  2. Taking care not to drip culture from the end of the pipette, place ten drops of culture to the centre of the plate or enough to cover a 5 pence piece..
  3. Replace lid of Petri dish.
  4. Place pipette in discard jar.
  5. Dip glass spreader in ethanol, flame and allow the ethanol to burn off.
  6. Lift the lid of the Petri dish to allow entry of spreader.
  7. Spread drops of liquid culture evenly around plate, rotating plate with left hand if this feels comfortable. Make sure the entire agar surface is covered.
  8. Replace lid of Petri dish.
  9. Flame spreader using ethanol.

- results of spread plate - Candida utilis- results of spread plate - Phaffia rhodozyma

image

  1. Lift a universal of sterile nutrient broth in the left hand.
  2. Remove the lid of the universal with the little finger of the right hand which still holds the charged loop or pipette. Do not put down the lid.
  3. Flame the neck of the universal.
  4. Insert the loop charged with inoculum into the sterile broth. Touch on the inside of the universal and withdraw.
  5. If using a pipette, gently release the required number of drops from the pipette into the culture and withdraw pipette.Do not agitate or cause bubbles in the liquid medium.Flame the neck of the universal, replace lid and place the universal on the bench.
  6. Flame the loop and place on heat resistant mat/ place pipette in discard jar.
  7. Tighten lid of universal to make secure but do not overtighten.

image 7

  1. Partially lift the lid of the Petri dish containing the sterile malt agar medium.
  2. Place the cube/bore of agar or fungal mycelium on to the centre of the sterile agar. Use a mounted needle which has been flamed in ethanol to assist with the transfer process if necessary.
  3. Withdraw implement.
  4. Dip the implement in ethanol, flame as above and place on heat resistant mat.