Cat. No. / ID: 338300
Expert designed assays allow you to realize the full potential of dPCR to best answer your specific biological questions. Partner with QIAGEN to design, execute and, optionally, verify your results, taking advantage of our custom assay design expertise.
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dPCR increases sensitivity in the detection and absolute quantification of rare events and sequence variants by creating a partitioning that increases the concentration of the specific target. For instance a mutation present at 0.1% can be detected in a background of wild-type genomic DNA using dPCR. The end-point amplification and detection over noise, with a binary positive/negative target calling, allows precise quantification, which is independent from amplification performance (affected, for example, by PCR inhibitors).
Based on your needs, the Custom dPCR LNA Assays are designed to work in a single- or multiplex setup, using an approach based either on a probe or on EvaGreen and a primer. Digital PCR provides the most sensitive and reliable method for detecting rare events over background. Custom dPCR LNA Assays use LNA-enhanced primers and probes, which increase assay specificity and sensitivity, making it possible to detect, for instance, a mutation present at 0.1% in a background of wild-type genomic DNA in a single nanoplate well.
To set up your dPCR experiment, add an aliquot of your genomic DNA sample (isolated from fresh, frozen or fixed samples) to the master mix included in the QIAcuity Probe PCR Kit or the QIAcuity EG PCR Kit along with your Custom dPCR LNA Assay. Each reaction is analyzed in one or multiple wells of a QIAcuity Nanoplate.
After loading and sealing the nanoplate, select the recommended cycling program on the QIAcuity instrument. Once the dPCR is complete, the number of copies/µl for each of your target sequences is shown in the QIAcuity Software Suite. You can find more details in the Custom dPCR LNA Assay Quick-start Protocol .