100x TAE buffer 5 blotting buffer for Northerns 
 

 
100x TAE5 is composed of 1 M Tris base, 50 mM Na₂EDTA, 500 mM NaOAc and approximately 43 mL of glacial acetic acid (to pH 7.8) per liter of 100x buffer.  
 
Autoclave or filter sterilize.  This buffer is used with the Hoeffer/LKB apparatus to transfer RNA to nitrocellulose or nylon.  The Hoeffer/LKB box requires almost 5 liters of 1x TAE₅.  Gels do not need to be presoaked to remove urea.  See the Northern protocol.  
Acetate is oxidized to carbonate during electrophoresis, raising pH.
 
50x Denhardt’s
 
 1%      5      gFicoll
 1%      5      gpolyvinylpyrolidone
 1%      5      gacetylated BSA
         x mL   Type I water
       500 mL  
 
Dissolve with mild heat and stir.  Spin 2000 rpm for 15 min.  Filter sterilize supernatant through 0.45 µ unit with 2 prefilters. 50x stock is difficult to filter sterilize unless prefilters are used.  Store aliquots at -20℃. Do not thaw at elevated temperatures.  
 
20x SSPE
 
  175.3 g           NaCl
  27.6 g             NaH₂PO₄
  7.4 g               Na2EDTA
  800 mL          Type I water 
  ≈20.5 mL     10 N NaOH to pH 7.4
  x mL               Type I water
  1000 mL  
 
Autoclave or filter sterilize.  SSPE can replace SSC plus NaPO₄, pH 6.7. 
 
50% Dextran sulfate MW 5000
 
                 2 g          Dextran sulfate MW 5000
                x mL       Type I water 
                4 mL  
 
Dextran sulfate MW 5000 works just as well as the traditional MW 500,000 material, but is significantly easier to dissolve.  Weigh dextran sulfate directly into anti-static treated tube (do not try to transfer from weighing paper or weigh boat).
Dextran sulfate supports the growth of bacteria.  Store at -20℃

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