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Applyo qPCR product for 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!
|Storage conditions||Store at 20°C. Short term storage at temperatures up to 30°C is also possible.|
|Shelf life||12 months|
|Concentration||Each Appylo qPCR Master-Mix Bead contains all components for 25 qPCR reactions à 25 µl.|
|Description||Applyo qPCR Beads contain all reagents for setting up a qPCR Master-Mix for detection of DNA in a lyophilized bead format. This includes among others a Hot-Start Taq DNA Polymerase and buffer in a seprate tube. The Master-Mix is designed for performing 25 qPCR reactions à 25 µL.|
|1 Tube with 1 Bead for 25 reactions||42,50 €|
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.
After reconstitution of the Master-Mix Bead the liquid Master-Mix is stable at -20°C, short time storage at 4°C is also possible. Avoid extended freeze-thaw cycles.
Resuspension of the Applyo Beads
Open the tube containing the Applyo Bead. Use 247.5 µL of supplied buffer for direct reconstitution 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|
|DNA sample||1 µl – 12.5 µl||20 – 100,000 copies / reaction|
|Nuclease-free water||Fill up to 25 µl|
Recommended cycling program:
|95°C||10 sec.||Repeat 45x*|