Preparation of Buffers
Traut’s Preparation
August 11, 2016
Present: Edward, Amir
What was achieved?
Prepared 45.4mL of 368.41uM of Traut’s reagent suspended in 5mM PBS/EDTA
Modifications to Protocol
- Transferred some Traut’s reagent from its original bottle to a 50mL Faclon tube, currently covered in foil and stored at 4°C
- added 45.4mL of 5mM EDTA/PBS to a 50mL falcon tube
- weighed 2.3 mg of Traut’s reagent into a microcentrifuge tube
- pipette 1mL of the EDTA/PBS into the microcentrifuge tube
- pipette the EDTA/PBS/Traut’s mixture back into the Falcon tube
- repeat 3x to wash the microcentrifuge tube
- aliquot the Traut’s solution into 20 microcentrifuge tubes, 1 mL each
- put the 20 tubes, and the Falcon tube in the –20 fridge
PBS Prep
August 25
- Made 500mL of 10X PBS by adding 2g KCl, 2.4g KH2PO4, 40g NaCl, 7.2 Na2HPO4
- first dissolved solids in ~300mL of MilliQ, adjusted pH to 7.2, then filled with MilliQ to 500mL
- After diluting the 10x PBS to 1X, make sure to check pH again. When I did it the pH increased by 0.8
- I think my original pH measurement was not accurate
- labelled and stored at 4°C
- used 50mL of the 10X PBS to make 500mL of 1X PBS