The drying conditions provided in the product manual are only an example of the tested cycles used in our labs. When air-drying, it is important to get a loss of weight (Loss on Drying, LoD), of approximately 70-75 %, which results in a dried gel containing only 5-10 % water.

Drying times may vary depending on the instrument used. When using this product for the first time with a specific oven, we recommend measuring the LoD using multiple tubes at regular times (e.g., after 60, 70, 80, 90 min) to define the best protocol and achieve a LoD of 74.5 ± 10.5 % or 70 ± 1 % based on the mix (with or without oligos) and the drying instrument used.

Temperatures can also be changed, but obviously lower temperatures will require longer times, while higher temperatures risk affecting the integrity of the mix components, so we recommend using 40 °C to minimise the chances of unexpected problems.

Although we successfully managed to lyophilise this mix, we do not recommend it be lyophilised. The excipient content has been optimised for air drying and this significantly reduces stability of the product lyophilisation. We recommend using this mix for air-drying only and recommend our Lyo-Ready product range for lyophilisation protocols.

Absolutely. This is our first universal All-in-One mix, meaning that it already contains the RTase. Critically, this does not affect the performance of our HS Taq polymerase, hence it can be used both with RNA or with DNA as template. We do not recommend simultaneous detection of RNA and DNA targets in a single multiplex reaction.

If 5 – 6 μL of mix (plus oligos) were dried in each well, it is sufficient to resuspend the gel with 20 μL of template in water. After resuspension, we recommend vortexing the sealed plate for about 1 min to achieve a homogeneous solution in each well.

Once dried, seal tubes/plates. In this way, they can be stored at room temperature for at least 1.5 year. For long-term storage, we recommend storing the plates inside zip lock bags with silica gel bags.

The dried mix behaves similarly to the wet mix, with no significant reduction of fluorescence or delay in amplification. It also shows the same sensitivity and efficiency as all our Probe 1-Step mixes, detecting down to 4 copies of viral genome.

We observed no significant loss of activity (i.e., no reduction of the fluorescence plateau and less than 1 Ct delay in amplification) when we incubated the dried gels at 37 °C for 3 months.

We successfully amplify 2 different sets of 4 targets simultaneously. External users report successfully running more complex multiplex reactions. Please share any feedback and results on this with us at [email protected].

On arrival the kit should be stored between -30 °C and -15 °C. If stored correctly the kit will retain full activity for 12 months

Please check that the instrument used to dry the mix is correctly set up, connected to a power source, and operating correctly.

Also be sure not to exceed a maximum of 5 – 6 μL of mix (including oligos and ROX dye) per well when drying down. Please ensure you are following the instructions provided in the manual. Optimal drying times may differ among different ovens. We recommend optimising the drying conditions for your reaction as necessary, starting with the indications provided in our manual.

We recommend using the smallest volume possible to minimize the time for the procedure. This means drying 5 μL of the 4x mix alone (for a 20 μL reaction). If primers, probes, and eventually ROX (if the PCR instrument requires it) are added before the process, stock solutions at high concentrations should be prepared, in order to add the smallest volume possible to the 4x mix. We recommend not exceeding 1 extra μL. Larger volumes would require longer drying times and higher LoD to reach the same percentage of remaining water in the dried product. When drying 6 μL of the mix with primers and probes, the LOD needs to be 74.5 % ± 0.5 %, and for that we recommend drying the mix at 40 °C for 80-90 min.

We do not recommend drying more than 5 – 6 μL of mix (plus oligos) per well following the instructions provided into the manual. Multiple plates can be dried at the same time for high throughput of manufacturing. We do not recommend air-drying larger volumes and recommend lyophilisation, using our Lyo-Ready range of products.