Multiplex Amplicon Quantification (MAQ)

MAQ is a method for the analysis of specific copy number variations (CNVs). The method consists of fluorescently labelled multiplex PCR with amplicons in the CNV (target amplicons) and amplicons with a stable copy number (control amplicons). After PCR, the fragments are size separated on a capillary sequencer. The ratios of target amplicons over control amplicons are calculated for the test sample and a reference sample. Comparison of these relative intensities results in a dosage quotient, indicating the copy number of the CNV in the test sample (Figure 1).

A MAQ analysis is very robust and extremely fast and easy to perform (Figure 2). The PCR amplicons are generated in a one-step, closed tube reaction, a warranty for easy upscaling .

We developed a software tool, MAQ-S, for the straightforward analysis of the generated chromatograms. The user imports the chromatogram files and selects the assay description file for the MAQ kit used. When the reference individuals of the experiment are indicated, the Dosage Quotients are automatically calculated and graphically represented in the Dosage Plot allowing simple visual verification.

Field of application:

• Confirmation of CGH data
• High throughput CNV analysis
• Screening for CNVs in candidate gene/region