Difference between revisions of "Loading a Protein Gel"

Latest revision as of 21:01, 18 July 2018

(Identify how much protein you will have to load in the wells (excel sheet))

1. Take the gel apparatus and place it onto the running adapter (positive and negative screws) make sure knobs from apparatus are facing out
2. Place the adapter into the running chamber and fill the inner chamber with running buffer

Running buffer 1x solution: 100 ml running buffer (on bench) + 900 ml DI water

1. Fill the outside of the chamber with running buffer (ensure there is two seperate areas of liquid)
2. Add 4 ul of loading buffer to 1.7 ml tubes, amount of tubes depends on how many samples you have
3. Place protein samples on ice (remember to keep them on ice the whole time)
4. Using the Precision plus (blue) ladder [kept in fridge door], pipette 3.6 ul into the first well on both gels, keep on ice
5. Vortex tube, pipette amount of protein needed into 1.7 ml tube containing loading buffer
6. Take up the liquid from the tube (sample + loading buffer) and carefully pipette this mix into the well next to the ladder.

To load: press pipette towards you against the larger glass and carefully go between the plates into the running buffer. Slowly dispense all of the liquid (should fall to the bottom of the well) and remove the tip. Pull any bubbles from the pipette up towards the surface so it does not disturb your sample

1. Repeat 7 & 8 until all protein samples are added
2. Fill any remaining lanes with loading buffer