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Illumina sequencing technology, sequencing by synthesis (SBS), is a widely adopted next-generation sequencing (NGS) technology worldwide, responsible for generating more than 90% of the world’s sequencing data. A fluorescently labeled reversible terminator is imaged as each dNTP is added, and then cleaved to allow incorporation of the next base. Since all 4 reversible terminator-bound dNTPs are present during each sequencing cycle, natural competition minimizes incorporation bias. The method virtually eliminates errors and missed calls associated with strings of repeated nucleotides (homopolymers). Illumina sequencing by synthesis technology supports both single-read and paired-end libraries. SBS technology offers a short-insert paired-end capability for high-resolution genome sequencing, as well as long-insert paired-end reads for efficient sequence assembly, de novo sequencing, and more. The combination of short inserts and longer reads increases the ability to fully characterize any genome.
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