How it Works

MassARRAY® with iSEQ™

The iSEQ application uses reference sequences as a comparative measure to identify microbes, viruses, or other haploid organisms with the discriminatory power down to a single nucleotide. An optimized protocol has been developed based on the MassCLEAVE™ chemistry to enable accurate identification.

The iSEQ assay is PCR based and allows for the amplification of a target region of interest. Following SAP treatment, the samples are subject to in vitro transcription and base-specific enzymatic cleavage of 500 – 800 bp PCR products of the target region of interest. The process is homogeneous and does not require purification of the PCR product or subsequent analytes. The workflow is automatable and thus amenable to high throughput. After sample conditioning, the cleavage mixture is resolved and data is acquired by the MassARRAY MALDI-TOF Mass Spectrometry. A protocol overview is provided in the figure below.

Key benefits of iSEQ process:

Sensitive

• Enables two levels of amplification, PCR and in vitro transcription

Cost- effective

• Miniaturization in 384 well format
• Requires low amount of starting material (2 - 5 ng DNA)

Accurate

• Offers 4 data points per amplicon