This protocols shows how to mix 0.5 M carbonate/bicarbonate buffer
|Item||Item article number||Distributor||Vendor|
|Sodium carbonate, anhydrous
|UltraPure™ DNase/RNase-Free Distilled Water||10977035||ThermoFisher||Invitrogen|
The carbonate/bicarbonate anion system has two pKa values, one at 6.4 and one at 10.3. Therefore, at pH 8-9, you will have both carbonate and bicarbonate present. One simple way to make your buffer is to make two 100mM solutions, one of NaHCO3 and one of Na2CO3. You can then mix various proportions of the two until you get the proper pH.
Make 0.5 M carbonate solution by adding 2.75 g in 50 mL nuclease-free water.
Make 0.5 M bicorbonate solution by adding 2.1 g in 50 mL nuclease-free water.
Combine 750 µL of carbonate solution into 23 mL of bicarbonate solution.
Check pH that it is 8.75.
Fill up to 50 mL with nuclease-free water.