Diluted PBS

Required Items:

• 25 ml Serological Pipette
• 500 mL volume metric flask
• empty 500 ml borosilicate glass bottle 

Stock solution:  (PBS - PHOSPHATE BUFFERED SALINE 10× concentrate - P5493) from sigma / Merck, Type II water

Preparing diluted PBS

Instruction:

1^st: Transfer 50 ml PBS stock solution into the empty 500 ml borosilicate glass bottle (work inside the bio LAF bench to avoid polluting the stock PBS solution)

2^nd: Transfer 450 ml type II water into the same above 500 ml borosilicate glass bottle. (work outside the LAF bench, on the lab bench)

3^rd: Autoclave the diluted PBS solution before using.

Work Steps

Step 1:  Use 75% antibac ethanol alcohol to spray the pack surface of 25 ml Serological Pipette, the surface of the 500 ml borosilicate glass bottle and the bottle of stock solution.

Step 2: right after spray each item and place them inside the bio LAF hood.

Step 3: Transfer 50 ml PBS stock solution into the empty 500 ml borosilicate glass bottle and close the cap, then take out the bottle from LAF.

Step 4: transfer 450 ml type II water with 500 mL volume metric flask into the 500 ml borosilicate glass bottle. (Work outside the LAF fume hood – just on lab bench).

Step 5: Autoclave the diluted PBS solution.