Bio-Rad Experion Protein Analysis Kits - Manual (Page 14)

4. Pipet 6 µl of each diluted sample into sample wells 1–10. Use the layout shown in
Figure 1.3.
S3 S2 S1 GS
S3 S2 S1 GS
S3 S2 S1 GS
S3 L GS G
Fig. 1.3. Chip layout for Test 1. S1–S3, sample numbers; GS, gel-stain solution; G, Pro260 gel; L,
Pro260 ladder.
5. Inspect all wells to make sure that there is no excessive bubble formation from pipetting.
Hold the chip above a light-colored background and look down through the wells
(Figure 1.4). Dislodge any bubbles at the bottom of the well with a clean pipet tip or by
removing and reloading the solution.
6. Pipet 6 µl diluted Pro260 ladder into the ladder well labeled L (Figure 1.3). Use the
Pro260 ladder within 8 hr of preparation. Every chip must have the Pro260 ladder
loaded into the ladder well labeled L.
Fig. 1.4. Bubble formation during loading of Experion Pro260 chips. Left, example of bubbles
trapped at the bottom of wells. The GS and G wells and sample wells 1, 3, and 4–6 contain no solution.
Well 10 is filled properly and has no bubbles; large bubbles have formed at the bottom of wells 8 and 9
and in the ladder well (L). Note the difference in the diameter of the light-colored circles in wells 10 and
L. Right, example of bubble formation at the surface of wells. Small bubbles have formed at the surface
of the three GS wells on the right side of the chip, and the rest of the wells have no bubbles. Surface
bubbles should not cause problems during a run, but bubbles at the bottoms of wells must be removed.
7. Place the loaded chip into the Experion electrophoresis station and start the run. It is
important to run a loaded chip immediately (within 5 min). Otherwise, excess
evaporation may occur, leading to poor results or to a chip performance error.
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