What is amplicon sequencing?

Amplicon sequencing is a targeted sequencing method that uses polymerase chain reaction (PCR) to selectively amplify specific DNA fragments or genomic regions of interest. Specific primers are designed to flank the target region and the resulting amplicons are then sequenced to determine the nucleotide sequence.

What is the difference between targeted sequencing and amplicon sequencing?

Targeted sequencing and amplicon sequencing both selectively sequence specific regions of the genome, rather than the whole genome. However, the difference between the two methods lies in how the regions of interest are selected and amplified.

Targeted sequencing amplifies through hybridization capture and uses probes or baits complimentary to the target region in order to selectively capture and enrich for specific genomic regions, such as genes, exons, and other functional regions of the genome. Targeted sequencing often offers greater coverage than amplicon sequencing, as the use of hybridization capture can help mitigate biases in the representation of certain genomic regions.

Amplicon sequencing amplifies DNA fragments using PCR. The target regions are defined by PCR primers designed to anneal specific sequences that flank the areas of interest, initiating the amplification process of the targeted DNA fragments. Amplicon sequencing is often more cost-effective than targeted sequencing as it does not involve the use of capture reagents.