Next-generation sequencing

Next-generation sequencing comprises high-throughput technologies that determine DNA sequences by reading millions of DNA molecules in parallel. Unlike Sanger sequencing, it produces large numbers of sequence reads in a single run.

DNA library preparation: The process of fragmenting DNA and adding platform-specific adapters to prepare it for sequencing. Adapters and fragments make DNA compatible with next-generation sequencing platforms.
DNA sequencing: The determination of the order of nucleotide bases in a DNA molecule. Next-generation sequencing is a high-throughput family within DNA sequencing.
Whole-genome sequencing: The sequencing of nearly all genomic DNA in an organism or sample. It uses high-throughput reads to survey coding and noncoding regions together.
Sanger sequencing: A DNA sequencing method that uses chain-terminating nucleotides to read individual DNA templates. Its lower throughput contrasts with the many-molecule parallelism of next-generation sequencing.
Human Genome Project: An international research effort that produced a reference sequence for the human genome. Its slower, large-scale sequencing effort preceded routine high-throughput genome reading.
Bridge amplification: A surface-based PCR method that creates clonal clusters of DNA fragments for sequencing. Illumina instruments amplify library fragments into clusters before detecting their bases.
Read depth: The number of sequencing reads covering a particular nucleotide position or genomic region. Depth indicates how many independent observations support a base call.
RNA-Seq: A method that sequences RNA-derived complementary DNA to measure and characterize transcripts. The same parallel sequencing platforms can profile gene expression and RNA variation.
Third-generation sequencing: Single-molecule sequencing technologies that read individual DNA molecules, often in real time. These platforms differ from many next-generation systems that sequence amplified clusters or templates.
Genome-wide association study: A study testing genetic variants across many people for statistical association with traits or disease. High-throughput genotyping and sequencing expanded the scale of variant discovery.

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