Sample Set:1. Two sets (1 set per b

Sample Set:
1. Two sets (1 set per blender jar) of ten (10) randomly selected spore strips removed
from self-contained biological indicators.
Procedure:
1. Dilution Scheme: Reference Section F (below) for suggested dilution plan. Adjust
dilutions as necessary to obtain plates in the range of 30 – 300 CFU per plate.
2. Place each set of 10 spore strips in a sterile 500 ml blender jar containing 100 ± 1.0
ml of sterile PBW (10-1 dilution). Label samples appropriately.
3. Immediately blend each set for 2 minutes ± 10 seconds. DO NOT ALLOW FIBERS
TO SETTLE OUT.
4. Immediately aseptically pipette 10.0 ml of the blenderized spore strips (10-1 dilution)
from each respective blender jar into a separate milk dilution bottle containing 90.0 ±
1.0 ml of sterile PBW (10-2 dilution). Insure that fibers in solution are uniformly
dispersed as sample is taken. Mix thoroughly by vortexing for at least 5 seconds.
5. Immediately pipette 1.0 ml of each respective 10-2 dilution into a separate test tube
containing 9.0 ± 0.1 ml sterile PBW (10-3 dilution); mix thoroughly by vortexing at least
5 seconds.
6. Pipette 1.0 ml of each respective 10-3 dilution into a separate test tube containing 9.0
± 0.1 ml sterile PBW (10-4 dilution); mix thoroughly by vortexing at least 5 seconds.
7. Pipette 1.0 ml of each respective 10-4 dilution into a separate test tube containing 9.0
± 0.1 ml sterile PBW (10-5 dilution); mix thoroughly by vortexing at least 5 seconds.
8. Pipette 1.0 ml of each respective 10-5 dilution into a separate test tube containing 9.0
± 0.1 ml sterile PBW (10-6 dilution); mix thoroughly by vortexing at least 5 seconds.
9. Prior to pipetting, revortex 10-4 dilution for at least 5 seconds. Pipette 0.5 ml of each
respective 10-4 dilution into a sterile, 100 x 15 mm, labeled Petri dish, in duplicate.
10. Prior to pipetting, revortex 10-5 dilution for at least 5 seconds. Pipette 1.0 ml of each
respective 10-5 dilution into a sterile, 100 x 15 mm, labeled Petri dish, in duplicate.