Custom dPCR LNA Assay (Probe)

Designed to order by QIAGEN's expert team using optimized internal design tools, this probe-based digital PCR assay is configured to match the specific requirements of each experiment. Each design is tailored to a target sequence, species, and application, and is available as a single or multiplex assay. Custom assays can be configured, executed, and optionally verified, supporting applications that include rare mutation detection, copy number variation, and NGS validation.

The assay uses LNA-enhanced primers and probes, which increase specificity and sensitivity and enable detection of variants present at 0.1% against a wild-type background in a single nanoplate well. Digital PCR achieves this level of sensitivity by partitioning the reaction, which increases the concentration of the specific target, and by using end-point amplification with binary positive/negative target calling for precise quantification that is independent of amplification performance.

To set up the experiment, a genomic DNA aliquot (isolated from fresh, frozen, or fixed samples) is combined with the master mix from the QIAcuity Probe PCR Kit along with the assay. Each reaction is analysed in one or multiple wells of a QIAcuity Nanoplate. After loading and sealing the nanoplate, the recommended cycling program is run on a QIAcuity instrument, and the number of copies/µl for each target sequence is displayed in the QIAcuity Software Suite.