|
|
| (10 intermediate revisions by 2 users not shown) |
| Line 1: |
Line 1: |
| ==Running SDS-PAGE==
| | [[category:Protein_Work]] |
| | |
| #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 and 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
| |