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This design is very different from that of Sanger sequencing—also known as capillary sequencing or first-generation sequencing—which is based on electrophoretic separation of chain-termination products produced in individual sequencing reactions. [6] This methodology allows sequencing to be completed on a larger scale. [7]
The workflow of a typical hybrid genome assembly experiment using second- and third-generation sequencing technologies. Figure adapted from Wang et al., 2012 [14]. One hybrid approach to genome assembly involves supplementing short, accurate second-generation sequencing data (i.e. from IonTorrent, Illumina or Roche 454) with long less accurate third-generation sequencing data (i.e. from PacBio ...
ChiRP-Seq is one of these new methods which uses the massively parallel sequencing capability of 2nd generation sequencers to catalog the binding sites of these novel RNA molecules on a genome. Although many have believed that RNAs mainly encode for proteins a very large portion of the eukaryotic genome is composed of RNAs that do not.
Together these were called the "next-generation" or "second-generation" sequencing (NGS) methods, in order to distinguish them from the earlier methods, including Sanger sequencing. In contrast to the first generation of sequencing, NGS technology is typically characterized by being highly scalable, allowing the entire genome to be sequenced at ...
2 Base Encoding, also called SOLiD (sequencing by oligonucleotide ligation and detection), is a next-generation sequencing technology developed by Applied Biosystems and has been commercially available since 2008. These technologies generate hundreds of thousands of small sequence reads at one time.
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These two methods were used to put together our human reference genome. However, since Sanger sequencing is low throughput and expensive, only a few genomes are assembled with Sanger sequencing. Second-generation sequencing reads are short, and these sequencing techniques can efficiently and cost-effectively sequence hundreds of millions of reads.
The advent of second-generation sequencing technologies has made it possible to obtain sequence information across the entire bacterial genome at relatively modest cost and effort, and MLST can now be assigned from whole-genome sequence information, rather than sequencing each locus separately as was the practice when MLST was first developed. [15]