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Microfluidic Sanger sequencing is a lab-on-a-chip application for DNA sequencing, in which the Sanger sequencing steps (thermal cycling, sample purification, and capillary electrophoresis) are integrated on a wafer-scale chip using nanoliter-scale sample volumes. This technology generates long and accurate sequence reads, while obviating many ...
Uses a short kmers to rapidly index genome; no size or scaffold count limit. Higher sensitivity and specificity than Burrows–Wheeler aligners, with similar or greater speed. Performs affine-transform-optimized global alignment, which is slower but more accurate than Smith-Waterman. Handles Illumina, 454, PacBio, Sanger, and Ion Torrent data.
DNA sequencing is the process of determining the nucleotide order of a given DNA fragment. So far, most DNA sequencing has been performed using the chain termination method developed by Frederick Sanger. This technique uses sequence-specific termination of a DNA synthesis reaction using modified nucleotide substrates.
In Sanger sequencing, one of the major problem regarding DNA polymerases is the discrimination against dideoxynucleotides, the chain-terminating nucleotides. Most of known DNA polymerases strongly discriminate against ddNTP; and thus, a high ratio of ddNTP to dNTP must be used for efficient chain-termination.
In genetics, shotgun sequencing is a method used for sequencing random DNA strands. It is named by analogy with the rapidly expanding, quasi-random shot grouping of a shotgun. The chain-termination method of DNA sequencing ("Sanger sequencing") can only be used for short DNA strands of 100 to 1000 base pairs.
Dideoxynucleotides are useful in the sequencing of DNA in combination with electrophoresis.A DNA sample that undergoes PCR (polymerase chain reaction) in a mixture containing all four deoxynucleotides and one dideoxynucleotide will produce strands of length equal to the position of each base of the type that complements the type having a dideoxynucleotide present.
In contrast to directed sequencing, shotgun sequencing of DNA is a more rapid sequencing strategy. [6] There is a technique from the "old time" of genome sequencing. The underlying method for sequencing is the Sanger chain termination method which can have read lengths between 100 and 1000 basepairs (depending on the instruments used).
Illumina and PacBio/Oxford Nanopore data, legacy 454 and Sanger data [17] 2011 / 2018 OS link: Newbler: genomes, ESTs 454, Sanger 454 Life Sciences: 2004/2012 C link: Phrap: genomes Sanger, 454, Solexa Green, P. 1994 / 2008 C / NC-A link: Plass Protein-level assembler: assembles six-frame-translated sequencing reads into protein sequences ...