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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 ...
The Sanger method became popular due to its increased efficiency and low radioactivity. The first automated DNA sequencer was the AB370A, introduced in 1986 by Applied Biosystems. The AB370A was able to sequence 96 samples simultaneously, 500 kilobases per day, and reaching read lengths up to 600 bases.
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).
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.
The method plots data points that represent a specific time and fluorescence intensity at various wavelengths of light to represent a DNA profile. [ 2 ] [ page needed ] In the field of genetics, an electropherogram is a plot of DNA fragment sizes, typically used for genotyping such as DNA sequencing . [ 3 ]
The Sanger method is limited and can produce a single read at the same time and is therefore suitable to generate DNA barcodes from substrates that contain only a single species. [30] Emerging technologies such as nanopore sequencing have resulted in the cost of DNA sequencing reducing from about USD 30,000 per megabyte in 2002 to about USD 0. ...
In the 1980s, low-throughput sequencing using the Sanger method was used to sequence random transcripts, producing expressed sequence tags (ESTs). [ 2 ] [ 14 ] [ 15 ] [ 16 ] The Sanger method of sequencing was predominant until the advent of high-throughput methods such as sequencing by synthesis (Solexa/Illumina).
The classical shotgun sequencing was based on the Sanger sequencing method: this was the most advanced technique for sequencing genomes from about 1995–2005. The shotgun strategy is still applied today, however using other sequencing technologies, such as short-read sequencing and long-read sequencing.