Consort EVS1000 Series Handmatig - Pagina 7
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buffer and thickness and percentage of gel. This will also affect quality of transfer so time course of
transfer should be performed for your particular samples and conditions.
When the blot time is completed, turn the power supply off.
Remove the cassettes from the main tank. Buffer can be reused but this may affect run quality if
continued.
Lift the hinge of each cassette and gently pry apart the blot sandwich and remove the membrane
from the gel.
The membrane is now ready to be probed.
Duration of blotting
One hour
Three hours
Buffer solutions for blotting
Do not adjust the pH when making these buffers as this will cause blot over-heating. The pH will vary
according to the freshness of the reagents used.
Towbin Buffer
•
25 mM Tris,
•
192 mM glycine,
•
20 % methanol pH 8.3
Towbin Buffer SDS
•
25 mM Tris
•
192 mM glycine
•
20 % methanol pH 8.3
•
0.05-0.1 % (w/v) SDS
Bjerrum and Schafer-Nielsen Buffer
•
48 mM Tris
•
39 mM glycine
•
20 % methanol pH 9.2
Dunn Buffer
•
10 mM NaHCO3
•
3 mM NaCO3
•
20 % methanol pH 9.9
EVS1100
EVS1200
100 V
100 V
400 mA
400 mA
50 V
50 V
200 mA
200 mA
EVS1000 Blot units
EV1300
100 V
400 mA
50 V
200 mA
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