Category:Protein expression: Difference between revisions

Revision as of 14:25, 17 February 2014

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
1. 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
1. The voltage increases to 200 V from 0 V
2. For 4% staking gel and 12.5 % separating gel the current reaches 50-60 mA and the power 10 -12 W
3. The current and the power decreases in time but not reach zero (if it reaches zero see the troubleshoot)
4. For the state type of gel it takes 35 - 40 min

The following 3 pages are in this category, out of 3 total.

@@ Line 1: / Line 1: @@
-This is a category for protein over-expression protocols.
+==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 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