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The page below is a sample from the LabCE course Precision Medicine-Molecular Mechanisms of Cancer Development and Actionable Genes. Access the complete course and earn ASCLS P.A.C.E.-approved continuing education credits by subscribing online.

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Next Generation Sequencing Technologies

There are many variations in the NGS technology. NGS applications often employ variations on PCR technology in order to produce enough genetic material to be analyzed. There are multiple technologies utilized in NGS applications. The two most common technologies are Illumina and Ion Torrent.
The Ion Torrent technology utilizes a sophisticated pH meter called 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 utilizes sequencing by synthesis (SBS). In this case the instrument can also detect new nucleotides as they are added to a growing DNA strand; however the technology used is different than that seen with Ion Torrent. Illumina uses fluorescently labeled bases and has 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.