RT-PCR & PCR Inhibitor Tolerance Screening Kit

The RT-PCR & PCR Inhibitor Tolerance Screening Kit contains five ready-to-use additives for systematic evaluation and optimization of PCR and RT-PCR assays. Depending on the assay, the additives improve tolerance to PCR inhibitors, allowing suitable reaction conditions to be identified for the respective enzyme system and sample matrix.

Systematic Screening with Five Additives

The five additives are supplied individually and tested separately. They can be evaluated with existing polymerase or reverse transcriptase systems, buffers and primer-probe combinations. Each additive is tested individually and compared with a control reaction without additive.

Improving PCR Inhibitor Tolerance

Inhibitors can affect the performance of PCR and RT-PCR assays. The five additives included in the Screening Kit allow their individual effects on a specific assay to be evaluated. Depending on the assay, enzyme system and sample matrix, individual additives may improve inhibitor tolerance.

Because their effects are assay-dependent, the additives are tested individually and compared with a no-additive control. This makes it possible to experimentally identify a suitable additive for the respective assay. The additive concentration can subsequently be further optimized for the specific application.

Screening Workflow and Evaluation

For initial screening, 2 µL of the selected additive is added to a 20 µL reaction, replacing an equivalent volume of water. Each additive is tested separately, together with a control without additive. All other reaction components and cycling conditions remain unchanged.

Results can be assessed using assay-relevant parameters such as Cq values, product yield, specificity and reproducibility. Further optimization of the additive concentration and validation in the intended sample matrix may be required depending on the application.

Assay-Dependent Optimization

The effect of each additive depends on the assay, enzyme system and sample matrix. The recommended additive volume is therefore intended for initial screening. The optimal concentration must be determined individually for each application.