Sequencing Is The Most Widely Used Targeted Sequencing Method

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Exome sequencing is the most widely used targeted sequencing method. The exome only represents less than 2% of the genetic code, but contains about 85% of known disease-related variants, making a cost-effective alternative to whole-genome sequencing2. Exome sequencing detects variants in coding exons, with the capability to expand targeted content to include untranslated regions and microRNA for a more comprehensive view of gene regulation3. DNA libraries can be prepared in one day and require only 4–5 Gb of sequencing per exome4. Exome sequencing enables researchers to focus their resources on the genes most likely to affect phenotype, it has a wide range of applications, including population genetics, genetic disease, and cancer studies3.

Like other next-generation sequencing methods, three general steps are required for exome sequencing: library creation, sequencing, and data analysis. The size and quality of tested samples are screened before establishing the library. In library creating, DNA molecules are fragmented into a suitable size and fused with platform-specific adapters. After size selection step free adapters elimination, PCR is performed to select for molecules containing adapters at both ends and to generate sufficient quantities for sequencing2. The sequencing is performed in specific machine such as Illumina Hi-Seq Platform, the working principles were discussed in detail in Topic Three, however, in exome sequencing, only exomes are selected, amplified and
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