iGEM 2019 IISER Kolkata

PREPARATION OF PHOSPHATE BUFFER SALINE

(PBS)

Sl.N Components Quantity

o.
1 NaCl 4g
2 KCl 0.1 g
3 KH2PO4 0.12 g
4 Na2HPO4 0.72 g
5 Autoclaved Distilled Water Up to 500mL
Total Volume 500 mL

Adjust the pH to 7.2 -7.4

PROCEDURE

● Take an autoclaved measuring cylinder, weigh all the 4

components, add them to the cylinder and dissolve in some
autoclaved distilled water using a magnetic stirrer.
● Adjust the pH to 7.2 and make the volume up to 500 mL with
autoclaved distilled water.
● Now take the PBS into the hood and step up the filter unit which
is autoclaved beforehand.
 iGEM 2019 IISER Kolkata

● The filter unit consists of a 0.22 um filter membrane and filtration

occurs only by suction with a vacuum pump.
● Add some PBS to the filtration unit and check for leaks, if there
are no leaks, start the vacuum pump and filter the media.
● Keep the filtered PBS at room temperature for 24 hours. If there is
any contamination, we can see some precipitation, if not, store
the media in a 4 °C fridge.