View source for Running SDS-PAGE

[[category:Gel]]
[[category:Protein_Preparation]]
[[Category:Protein expression]]

[[:File:SDS_PAGE.pdf|SDS PAGE]]

==Running SDS-PAGE==

#Make ready the samples to run
#Make ready the sample running buffer
#Make ready the gel
#Have the SDS-PAGE apparatus
#Place the the gel in the SDS-PAGE stand
#Place the stand with the gel in the SDS-PAGE apparatus bath
#fill the space between the gels with running buffer
#Take off the combs of the gel (take it out gently)
#Load the sample to each well
#Do not use the wells at the left and right end of the gel 
#fill the SDS-PAGE apparatus bath with running buffer to the bottom of the gel
#Place the cover of the SDS-PAGE 
## '''N.B''' Make sure the correct plug is connected to the correct electrode (red goes to red and black goes to black)
#Plug the cables to the SDS-PAGE apparatus power source
#Turn of the SDS-PAGE power source
#Set to constant voltage
#Use 200 V 
#Press the Run button to start the electrophoresis
## The voltage increases to 200 V from 0 V
##For 4% staking gel and 12.5 % separating gel the current reaches 50-60 mA and the power 10 -12 W
##The current the the power decreases in time but not reach zero (if it reaches zero see the troubleshoot)
##For the state type of gel it takes 35 - 40 min

===Troubleshoot===
#The power drops to zero
* really old buffer
* the space in between the gels is not filled with  buffer to the top