Running A Development Method - GE PhastSystem User Manual

5. Operation
5.6 Running a
development
method
54
The procedure for running development methods comprises four steps:
making up the solutions, connecting the bottles to the correct ports with
the PVC tubing, inserting the gels and pressing the start key. The rest is
automatic. The procedure is described below.
Preparing the development unit
You should always keep a fresh stock of solutions. Filter solutions to
keep the channels in the development unit clear and to avoid
precipitation on the gel(s). We recommend that you label the bottles
and the tubing (use the yellow tubing markers to mark the tubes)
with their corresponding port number.
1. Remove the caps on the cap set from the ports that you plan to
use.
2. Connect the ports (1-9) as required to the solution bottles with
PVC tubing.
3. Connect port 0 to waste: Use an empty bottle.
4. Check for kinks in the tubing. Make sure the tubing is securely
submerged in the solutions.
Important! The chamber fills with approximately 70 ml of
solution. The bottles should be filled with at least
75 to 80 ml of solution to allow for the residual
solution in the tubing.
5. Open the lid of the development chamber by pressing on the
right end of the red bar.
6. Check that the chamber gasket (on the lid) is secure.
Inserting the gels
7. Remove one gel from the separation bed with a pair of forceps
(use the tab of the gel backing). Be careful not to touch the gel
surface with your fingers since fingerprints stain and cloud the
protein band.
8. Slide the gel, gel surface down, into the upper position of the
gel holder.
Remove the other gel and slide it, gel surface up, into the lower
position of the gel holder.
Note! If you are developing only one gel, slide it into the lower
position, gel surface up.