Bio-Rad Peelable Foil Heat Seal - Manual (Page 12)

6
Chapter 2 — Using the PX1 PCR Plate Sealer
Optimizing Heat Sealing
Sealing optimization is a crucial step to ensure a
high-quality seal. Sealing a plate for too long or at
too high a temperature can result in oversealing and
potentially affect the quality of data in optical assays,
including qPCR. Undersealing a plate may permit
sample evaporation.
Tip: Optimize sealing conditions using an empty plate.
1. Seal an empty plate with the recommended
sealing parameters listed in Chapter 5.
2. Carefully examine the entire seal and plate surface.
Look for even sealing around each individual well
and across the entire plate (Figure 9).
3. Make sure well rims are not significantly deformed.
Note: Minimizing well deformation is especially
important for qPCR assays. Significant well
deformation could affect optical readings.
4. If using a peelable seal, peel off the seal and
examine the adhesive patterns where the wells
contacted the seal. Look for uniform circular
imprints across the entire seal (Figure 10).
5. Optional: fill a sample plate with a colored liquid
such as 0.01% bromophenol blue solution.
Examine sample volumes to ensure pipetting
accuracy. Seal the plate. Thermal cycle the plate
and examine all wells for signs of evaporation
(Figure 11).
6. Save optimized sealing protocol so that it can
be easily accessed for future experiments. Refer
to Storing and Using Sealing Protocols section for
detailed instructions.
Fig. 10. Uniform and complete
circles demonstrate good sealing.
Fig. 9. Impressions should have
uniform width around individual
wells and across entire plate.
Fig. 11. Examine sample volumes
after cycling. In this demo,
corner wells were poorly sealed to
demonstrate sample evaporation.