HighPoint/CannonLab:Buffer and Osmolyte Solutions

<owwmenu font="arial, helvetica, sans-serif" bold="1"

       color="white" bgcolor="indigo" hovercolor="white"
       bghovercolor="gray" topfontsize="10" fontSize="8" pagewidth="750"
       image="CannonLabIMG01.JPG" lab="MGSC/CannonLab">
   Home=Lab-Home
   Lab Members=#, People=People, Contact=Contact Information, Associates=Collaborators and Former Members
   Research=#,Protein Membrane=Protein Membrane Interactions, Osmolytes=Osmolyte Effects
   Protocols=#, Vesicles=Lipid Vesicle Preparation, Solutions=Buffer and Osmolyte Solutions, Fluorescence=Fluorescence Spectrometry, UV-Vis=UV-Vis Spectrometry, Pipetting=Pipetting Exercise
   Notebooks=#, Master List=Notebook, Dr Cannon=Notebook/Jonathan Cannon
   Links=#, Courses=Course Pages

</owwmenu>

Preparing Buffers

Always wear gloves to avoid contamination of your solutions with bacteria!

Stock Solutions

1. Prepare stock solutions of buffer components (typically 1 M)
2. Rinse clean storage bottle and filter flask an extra time
   • Use type I, 18 MΩ purified water
   • Drain and use your best aseptic technique from this point on
3. Filter stock solutions through 0.2 μm filter
4. Store in bottle for future use
   • Keep the bottle closed as much as possible
   • Always wear gloves and use clean instruments for removing aliquots
   • This will extend the life of your stock solution before bacteria begin to grow in it

Filtering

Nalgene Filter Assembly

Filter Flask

• The filtration assembly is put together as in the image on the left
  1. Place a 45 mm diameter, 0.2 μm pore size filter between the top and bottom halves
  2. Close with the metal clamp
  3. Place the entire assembly onto the mouth of a filter flask
  4. Attach the side arm to an aspirator
  5. Fill the reservoir with the solution to be filtered
• Repeat step 5 until the entire solution is filtered
• If the speed of filtration reduces drastically, consider replacing the filter membrane