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RT-qPCR Master-Mix Bead for 25 Reactions

Applyo RT-qPCR product for reverse transcriptase – quantitvative PCR analyses with target detection possible in 5-plex (e.g. detection with 488 nm, 533 nm, 552 nm, 581 nm, 652 nm, 678 nm wave length depending on the filter setup of your thermocycler).


For in vitro use and RUO only!

Applyo Beads
Shipping Room temperature
Storage conditions Store at 20°C. Short term storage at temperatures up to 30°C is also possible.
Shelf life 12 months
Form Lyophilized
Concentration Each Appylo RT-qPCR Master-Mix Bead contains all components for 25 RT-qPCR reactions à 25 µl.
Description Applyo RT-qPCR Master-Mix Beads contain all reagents for setting up a RT-qPCR Master-Mix for detection of RNA (and DNA) in a lyophilized bead format. This includes among others a Hot-Start Taq DNA Polymerase and reverse transcriptase and buffer in a separate tube. The Master-Mix is designed for performing 25 RT-qPCR reactions à 25 µL.

Available Options

1 Tube with 1 Bead for 25 reactions


Applyo RT-qPCR Beads

Applyo Beads are stable under ambient conditions. Exposure to high moisture (>80 % rH) for more than 30 minutes or placing beads on ice may result in moisture condensation and may impair the PCR-performance due to the reduced solubility of the Applyo Beads. Therefore, limit the exposure to humidity (e.g. in the air) to a minimum until the bead is reconstituted with the supplied buffer.

Resuspended Master-Mix made from Applyo RT-qPCR Master-Mix Bead for 25 Reactions

Applyo Liquid Master-Mix is stable at -20°C, short time storage at 4°C is also possible. Avoid extended freeze-thaw cycles.

Recommended RT-qPCR protocol for Applyo RT-qPCR Beads

Handling of the Applyo Beads

Open the tube containing the Applyo Bead. Use 247.5 µL of supplied buffer for direct resuspension in the tube the Applyo Bead is provided in. Mix and centrifuge Master-Mix down gently. The Master-Mix is now 2x concentrated. For each 25 µL reaction use 12.5 µL of the 2x Master-Mix and fill-up to 25 µL with Primer, Probes and target diluted in PCR-water or 10 mM Tris-buffer.

Protocol at a glance

Component Stock conc. Volume in 25 µl assay Final conc.
Resuspended 2x Master-Mix 2x 12.5 µl 1x
Primer 1 µl – 12.5 µl ~ 200 – 800 nM
Probes 1 µl – 12.5 µl ~ 50 nM – 200 nM
RNA sample 1 µl – 12.5 µl 20 – 100,000 copies / reaction
Nuclease-free water Fill up to 25 µl

Recommended cycling program:

Temperature* Time*
50°C 10 min.
95°C 1 min.
95°C 10 sec. Repeat 45x*
58°C 10 sec.
72°C 20 sec.
Signal readout