Diluted PBS

Required equipment

• 25 mL Serological Pipette
• 500 mL volume metric flask
• 500 mL borosilicate glass bottle
• PBS (PHOSPHATE BUFFERED SALINE 10× concentrate - P5493 from sigma / Merck)
• Type II water

Preparing diluted PBS protocol

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.
2. Right after spray each item and place them inside the bio LAF hood.
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. Work inside the LAF bench to avoid polluting the stock PBS.
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 – on the lab bench.
5. Autoclave the diluted PBS solution before use.

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.