User:Matt Hartings/Notebook/AU Biomaterials Design Lab/2011/10/25
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Today we are going to be running a protein gel to determine how well we have been expressing and purifying the MBP-intein fusion. (Groups 2 and 3 have thus far done the expression and purification).
We will be using the Bio Rad Mini Protean system. The instruction manual can be found here. Each group will have to cast its own gel. We will be making a discontinuous polyacrylamide gel (so follow those instructions). We will be making a 12% gel. Each group will need 10 mLs of each gel solution for casting. From there, follow along the directions.
Resolving gel solution for a 12% discontinuous gel (per 10 ml)
Stacking gel solution for the gel we'll run (per 5mL)
Because we need to make lots of solutions, each group will be responsible for putting together 1 or more solutions. Please pay attention to what you need to make and begin making these buffers and solutions IMMEDIATELY upon entering lab. Everyone will be relying on you for quick and efficient preparation of buffers. This lab will have multiple steps (prepping gels, casting running layer, casting the resolving gel, casting the stacking gel, loading protein samples, running the electrophoresis, staining the gel, rinsing the stained gel). Please be efficient with your preparation! The best way to do this is to be prepared coming into class.
Again, be quick. Be courteous.
Once we have cast our gels. Tamra will go over some proper methods for loading and running the SDS-PAGE. After we run the gels, we will stain them and rinse the stain overnight.
This area is for any observations or conclusions that you would like to note.