There are many variations of NGS technology. NGS applications often employ variations on PCR technology in order to produce enough genetic material to be analyzed. There are several vendor-specific technologies utilized in NGS applications. The two most common technologies are Illumina and Ion Torrent.
The Ion Torrent technology utilizes a sophisticated pH meter (a semiconductor) to determine when a nucleotide has been incorporated into a growing DNA strand. It does this in hundreds of individual wells so that numerous reactions can occur simultaneously.
The Illumina technology performs sequencing by synthesis (SBS). In this case, the instrument detects new nucleotides as they are added to a growing DNA strand by using fluorescently-labeled bases with a laser to determine which base was added as it occurs.
NGS has much higher throughput than Sanger sequencing, meaning more patients can be analyzed per run. This is true because NGS often utilizes numerous primers in the same reaction to create hundreds of different PCR products covering the different targeted regions. Furthermore, individual patient samples can be barcoded with a specific sequence allowing numerous patient samples to be combined into a single tube. The data analysis at the back-end of NGS will allow the separation of the data from each patient.
