Whole exome sequencing (WES) is a targeted next generation sequencing method that identifies all the protein-coding genes (exons) in the genome. Regions of interest are hybridized to target-specific, biotinylated oligos and separated from rest of the DNA. By enriching for exons, you can focus on genomic regions relevant to your study.
This detailed overview walks you through major advances in capture and enrichment technology, types of targeted next generation sequencing, their applications, and more.
Exome sequencing is a type of targeted next generation sequencing. After genomic material is extracted from the sample, libraries must be prepared. Library prep includes the addition of adapters to identify the samples or molecules in the sample and to help the DNA or RNA adhere to the sequencing apparatus. Exome sequencing specifically enriches or captures the exome before the sequencing step.
The xGen Exome Research Panel consists of 5′ biotin–modified oligonucleotide probes that are individually synthesized and individually analyzed by electrospray ionization mass spectrometry (ESI-MS) and optical density (OD) measurement. Individual probes are made in large lots and then aliquoted to maintain reproducibility. The probes are then normalized before pooling to ensure that each probe is represented in the panel at the correct concentration. Probes that fail quality control are resynthesized. This rigorous manufacturing process gives the xGen Exome Research Panel a unique advantage over array-derived pools, in which missing or truncated probes cannot be identified before sequencing. Using IDT proprietary synthesis methods, even probes with high GC and AT content are appropriately represented in the panel.
To provide increased depth of coverage and enable high multiplexing of samples, the xGen Exome Research Panel targets only the coding sequences (CDS) of human coding genes in the RefSeq database.
Working in an area that would benefit from exome sequencing? Just starting? See how you can easily improve your workflows and results.
Learn how other scientists have used exome sequencing technology in the field.
Read our articles about whole genome sequencing.