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The company's first scientific instrument, called the PacBio RS, was released to a limited set of customers in late 2010., with full commercial release in early 2011. [17] [8] Sequencing provider GATC Biotech was selected by Pacific Biosciences as its first European service provider in late 2010. [18]
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 ...
Sequencing technologies vary in the length of reads produced. Reads of length 20-40 base pairs (bp) are referred to as ultra-short. [2] Typical sequencers produce read lengths in the range of 100-500 bp. [3] However, Pacific Biosciences platforms produce read lengths of approximately 1500 bp. [4] Read length is a factor which can affect the results of biological studies. [5]
In bioinformatics, sequence assembly refers to aligning and merging fragments from a longer DNA sequence in order to reconstruct the original sequence. [1] This is needed as DNA sequencing technology might not be able to 'read' whole genomes in one go, but rather reads small pieces of between 20 and 30,000 bases, depending on the technology used. [1]
Computational genomics refers to the use of computational and statistical analysis to decipher biology from genome sequences and related data, [1] including both DNA and RNA sequence as well as other "post-genomic" data (i.e., experimental data obtained with technologies that require the genome sequence, such as genomic DNA microarrays).
Assembly of single cell data is difficult due to non-uniform read coverage, variation in insert length, high levels of sequencing errors and chimeric reads. [ 8 ] [ 12 ] [ 13 ] Therefore, the new algorithmic approach, SPAdes, was designed to address these issues.
Experimental design is further complicated by sequencing technologies with a limited output range, the variable efficiency of sequence creation, and variable sequence quality. Added to those considerations is that every species has a different number of genes and therefore requires a tailored sequence yield for an effective transcriptome.
The application of massively parallel sequencing also greatly increases the amount of sequence data generated, which require high-throughput bioinformatic analysis pipelines. [90] The sequence-driven approach to screening is limited by the breadth and accuracy of gene functions present in public sequence databases.