Next-Generation Sequencing, a high resolution, sensitive and cost-effective solution method for multiple gene analysis

• The capture technique involves the use of specific probes or baits to capture the target sequences from the DNA sample. Then, the captured DNA fragments are sequenced using NGS technology. Capture sequencing allows for the sequencing of multiple genes simultaneously. This technique is well suited for sequencing a large number of genes or exons.

• The amplicon technique, on the other hand, uses PCR (polymerase chain reaction) to amplify the target regions of interest from the DNA sample. The PCR primers are designed to flank the target regions, and the amplified fragments are sequenced using NGS technology. Amplicon sequencing is typically employed for targeted sequencing of smaller regions, or a small panel of genes >500 kb. It can be challenging to design PCR primers that can efficiently amplify all regions of interest. In addition, the complexity of the PCR reaction can increase with the number of regions being targeted, which can lead to issues such as primer-dimer formation and non-specific amplifications.

For assay development, amplicon-based methods have been preferentially used for their shorter preparation time and small DNA input amounts. However, capture sequencing has emerged as an alternative approach because of its higher testing accuracy and reduced susceptibility to artifacts.