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

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(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...)
 
(Filtering)
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===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''
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* ''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 12:34, 23 February 2012

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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