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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 ...
Because of its comparative ease, the Sanger method was soon automated and was the method used in the first generation of DNA sequencers. Sanger sequencing is the method which prevailed from the 1980s until the mid-2000s.
The first DNA sequencing methods were developed by Gilbert (1973) [8] and Sanger (1975). [9] Gilbert introduced a sequencing method based on chemical modification of DNA followed by cleavage at specific bases whereas Sanger's technique is based on dideoxynucleotide chain termination. The Sanger method became popular due to its increased ...
Sanger's approach was described in 2001 as one of the two fundamental methods for sequencing DNA fragments [1] (the other being the Maxam–Gilbert method [5]) but the Sanger method is both the "most widely used and the method used by most automated DNA sequencers."
The sequencing occurs for millions of clusters at once, and each cluster has ~1,000 identical copies of a DNA insert. [12] The sequence data is analyzed by finding fragments with overlapping areas, called contigs, and lining them up. If a reference sequence is known, the contigs are then compared to it for variant identification.
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.
The method used in this study, which is called the “Sanger method” or Sanger sequencing, was a milestone in sequencing long strand molecules such as DNA. This method was eventually used in the human genome project. [5] According to Michael Levitt, sequence analysis was born in the period from 1969 to 1977. [6]
Although Maxam and Gilbert published their chemical sequencing method two years after Frederick Sanger and Alan Coulson published their work on plus-minus sequencing, [2] [3] Maxam–Gilbert sequencing rapidly became more popular, since purified DNA could be used directly, while the initial Sanger method required that each read start be cloned for production of single-stranded DNA.