Heat sealing films and foils efficiently seal microplates for standard, real-time, and digital PCR. These sealing solutions deliver reliable data by minimizing sample evaporation during thermal cycling.
Integrating the PX1 PCR Plate Sealer into the Thermal Cycling Workflow Helps Generate Highly Reproducible and Uniform PCR Data
Real-time PCR experiments using the PX1 PCR plate sealer in the workflow demonstrate highly uniform amplification curves. A, a SYBR® Green uniformity assay examined amplification uniformity across an entire 96-well plate sealed using the PX1 PCR plate sealer. The extremely low quantification cycle (Cq) standard deviation of all 96 wells (0.09 cycles) demonstrates exceptional experimental uniformity. B, the SYBR® Green uniformity melt curve analysis demonstrates a single, consistent melt peak across all wells, which indicates product specificity is maintained in plates sealed with the PX1 PCR plate sealer. RFU, relative fluorescence units.
The optically clear heat seal prevents sample evaporation that can lead to large Cq standard deviations. Five-color multiplex linearity test demonstrates highly reliable qPCR results. A–E, fluorescence data from a series of tenfold dilutions of plasmid DNA (108–102 copies) amplified using reporter dyes to monitor five targets. The standard curves of five multiplexed genes have an R2 >0.990. F, standard curves generated from data in A–E. Reaction efficiencies are between 90 and 105%. Cq, quantification cycle; RFU, relative fluorescence units.
Which heat seal is best for my PCR application? Use the Heat Sealing Films and Foils table to determine the seal that best suits your experimental needs.
How do I determine the best sealing time and temperature? Optimal sealing parameters vary according to the heat films and microplates used. We recommend sealing an empty plate using the guidelines found in Suggested Heat Sealing Parameters. After sealing, examine the entire plate for effective sealing and optimize sealing conditions if necessary.
Some of my wells are not fully sealed. What should I do? Sealing temperature and/or sealing time may be increased to help improve sealing performance. Use an empty plate to optimize seal conditions. See the instruction manual for additional details about seal optimization.
When should I use the sealing frame? The sealing frame helps secure the heat seal in place and prevents the film from moving during sealing. The sealing frame must be used for the permanent clear heat seal and the peelable foil heat seal.
What happens if I accidently place a seal upside down when sealing? The PX1 heated platen was designed with an easy to clean, nonstick surface. See the PX1™ PCR Plate Sealer Instruction Manual for directions on cleaning the heated platen. Ensure the PX1 sealer cools for at least 1 hour before cleaning.
Is it safe to leave the heat sealer on when unattended? Yes. The PX1 PCR plate sealer was designed for reliability and safety. The instrument has an adjustable auto-off feature that automatically shuts the heater off when not in use for a designated amount of time. Simply touch the screen and the instrument will automatically resume heating.
Can I reseal the same plate twice? A plate sealed with a peelable film may be resealed after peeling an existing film. However, resealing a plate for subsequent use in real-time PCR is not recommended. A plate sealed with a pierceable foil heat seal may also be resealed by sealing another pierceable foil heat seal directly on top. Note: only a single seal can be used in the QX100™ Droplet Digital™ PCR system workflow. Sealing parameters may need to be adjusted.
The current platen temperature is higher than the required sealing temperature. How can I reduce the time needed to reach sealing temperature? The heated platen cools passively. Opening the PX1 PCR plate sealer drawer can help accelerate the cooling process.
Can I use traditional adhesive or pressure-sensitive seals with the PX1 sealer? No. Our heat sealing films feature a special hot melt adhesive that effectively seals microplates when used with the PX1 sealer. The temperature and pressure delivered by the PX1 will not successfully seal a microplate when traditional adhesive and pressure-sensitive seals are used.
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