It involves the use of PCR to create multiple copies of a targeted DNA called amplicons. An amplicon is the product of PCR amplification. Amplicon sequencing is one of the most regularly carried out processes in molecular biology. Unlike the traditional PCR that employs two PCR primers, the amplicon sequencing panel utilizes multiple sets of PCR primers (multiplex PCR). This results in copying multiple regions of the genome simultaneously. Libraries are created by ligating the amplicons to adapters. This is followed by sequencing using next generation sequencing methods. Amplicons from different samples can be pooled and a barcode is added to samples for identification using analysis.

Amplicon sequencing enables the screening for many genetic variants, provides depth coverage of the targeted sequence, and allows sequencing of difficult targets like GC-rich regions. Amplicon sequencing is used for the identification and characterization of variations, sequencing microbial 16S rRNA gene from multiple species in metagenomics samples.

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