C.B.S. Scientific HTLE-7002 - Manual (Page 14)

C. B. S. œ Scientific 14 HTLE-7002
A. Choice and Preparation of Electrophoresis Buffers
Listed below are recipes for the most commonly used buffers. Use reagent quality solvents
and deionized water at all times. Pyridine is particularly unstable and should not be used if it
is yellow in color. Buffers can be prepared well in advance of electrophoresis if in containers
with air tight lids. Check the pH of solutions after they are prepared. The actual pH of the
buffer may be slightly different depending on the grade of solvents or the type of deionized
water used. For example, the pH 1.9 buffer we use is sometimes actually pH1.8. If your
solution is off by more than a pH unit then check to see that the correct dilution of each
component was added and that your pH meter is properly calibrated. Do not adjust the pH of
the buffers by using concentrated acid or base. Its best to keep a diary of the pH of buffers
as they are prepared.
For 2.01 of each buffer:
pH 1.9 Buffer
formic acid (88%)
50ml
glacial acetic acid 156ml
deionized water 1794ml
pH 3.5 Buffer
glacial acetic acid
100ml
pyridine 10ml
deionized water 1890ml
pH4.72 Buffer
n-butanol
100ml
pyridine 50ml
glacial acetic acid 50ml
deionized water 1800ml
pH 6.5 Buffer
glacial acetic acid
8ml
pyridine 200ml
deionized water 1792ml
pH 8.9 Buffer
ammonium carbonate
20g
Deionized water 2000ml