HighPoint/CannonLab:Buffer and Osmolyte Solutions: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(New page: {{CannonLabMenu}} ==Preparing Buffers== ===Stock Solutions=== # Prepare stock solutions of buffer components (typically 1 M) # Rinse ''clean'' storage bottle and filter flask an ''extra...)
 
Line 16: Line 16:


===Filtering===
===Filtering===
[[Image:FilterSupport.jpg|thumb|left|Nalgene Filter Assembly]]
[[Image:VacuumFilterFlask.jpg|thumb|Filter Flask]]
* ''The filtration assembly is put together as in the image on the left''
*# Place a 45 mm diameter, 0.2 μm pore size filter between the top and bottom halves
*# Close with the metal clamp
*# Place the entire assembly onto the mouth of a filter flask
*# Attach the side arm to an aspirator
*# 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''


===General rules for solution prep===
===General rules for solution prep===

Revision as of 13:34, 23 February 2012

<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

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

General rules for solution prep

Adjusting pH

Useful Recipes

1 M K2HPO4

1 M KH2PO4

1 M NaAcetate

1 M Acetic Acid

pH 7 buffer, 10 mM Phosphate, 50 mM NaCl

Retrieved from "https://openwetware.org/mediawiki/index.php?title=HighPoint/CannonLab:Buffer_and_Osmolyte_Solutions&oldid=587315"