Quantify Nucleic Acids with Unparallelled Accuracy and Precision

Quantify Nucleic Acids with Unparallelled Accuracy and Precision

Next Evolution in Nucleic Acids Quantification

Unlike traditional qPCR which only enables relative quantification, Combinati’s emulsion-free digital PCR (dPCR) platform provides absolute quantification of targets without the need for standard curves. The Absolute Q utilizes Microfluidic Array Partitioning (MAP) technology to break up a bulk reaction into tens of thousands of individual reactions enabling each target to be counted precisely. Accurate quantification is important for applications such as determining CRISPR editing efficiency or concentration determination of next generation sequencing libraries, but is also essential for applications where detecting small variances with higher precision is important- such as gene expression analysis.
Coupling a workflow that’s designed to be just like that of qPCR with 4-color multiplexing capability, Combinati’s Absolute Q allows users to easily transition current qPCR assay to dPCR without the need to adapt sample preparation steps or assays – saving time and improving data quality. .

From Oligo Design to Digital PCR: Multi-Color Assays with |Confidence|

IDT and Combinati come together to show you how simple it can be to make the transition from qPCR to dPCR. Designing a custom 4-plex assay can be easier than you think!
During this webinar our speakers cover assay development for gene expression and copy number variation applications. This talk is particularly useful for scientists who are looking for flexible and customizable digital PCR solutions as well as for scientists looking to make the switch from qPCR to dPCR.

Precision measurement of CRISPR Gene Edits

Digital PCR is aptly suited for the analysis of genome editing applications, such as CRISPR/Cas9 mediated knock-ins and knock-outs. This is largely enabled by dPCR’s fundamental principle of absolute quantification, which provides quantification of nucleic acid targets without the need for orthogonal standard curves. This method of quantification is more consistent and more accurate, particularly for rare or low concentration targets. Using two separate assays, this application note highlights how the Absolute Q digital PCR platform can be used to study the efficiency of CRISPR mediated genome edits

4-color digital PCR discrimination of a quad-allele SNP

To demonstrate 4-color optical multiplexing for single nucleotide difference discrimination, a 4-plex assay was designed in collaboration with Integrated DNA Technologies for a set of alleles in the CYP2C19 gene (rs12248560). The cytochrome P450 enzyme mediates the primary metabolism of many drugs. Polymorphisms in this gene alter metabolism of certain drug compounds. The polymorphism rs12248560, an ultra-fast metabolism phenotype, has been linked to more favorable outcomes for breast cancer patients receiving the drug tamoxifen.

Quantification of Illumina NGS Libraries on the Absolute Q

In order to maximize next generation sequencing (NGS) runs, the loading concentration of NGS libraries must be carefully controlled. Both over- and under-loading will decrease the amount of usable reads by limiting the number of sequenceable clusters. Library quantification using digital PCR (dPCR) is commonly used for accurate concentration determinations of complete and sequenceable library fragments.