QuantStudio System qPCR and dPCR Bundle

Digital PCR

Real-time qPCR

Overview

Measures the fraction of negative microreactions to determine absolute copies.

Measures PCR amplification as it occurs in a bulk reaction mix.

Quantitative?

Yes. The fraction of negative microreactions is fit to a Poisson statistical algorithm.

Yes. Data is collected during the exponential growth phase of PCR when the quantity of the PCR product is directly proportional to the amount of template nucleic acid.

Applications

• Absolute quantification of viral load
• Absolute quantification of nucleic acid standards
• Absolute quantification of next-gen sequencing Libraries
• Rare allele detection
• Absolute quantification of gene expression
• Analysis of multiple targets on a single molecule through multiplexing

• Quantitation of gene expression
• Microarray verification
• Quality control and assay validation
• Pathogen detection
• SNP genotyping
• Copy number variation
• MicroRNA analysis
• Viral quantitation
• • siRNA/RNAi experiments

Advantages

• No need to rely on references or standards
• Desired precision can be achieved by increasing total number of PCR replicates
• More tolerant to some PCR inhibitors
• Capable of analyzing complex mixtures
• Provides a linear response to the number of copies present to allow for small fold change differences to be detected

• Increased dynamic range of detection
• No post-PCR processing
• Detection is capable down to a 2-fold change
• Collects data in the exponential growth phase of PCR
• An increase in reporter fluorescent signal is directly proportional to the number of amplicons generated
• The cleaved probe provides a permanent record amplification of an amplicon