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A DNA microarray is a device for high-throughput investigations widely used in molecular biology and in medicine. It consists of an arrayed series of microscopic spots (‘features’ or ‘locations’) containing few picomoles of oligonucleotides carrying a specific DNA sequence.
[76] [77] [78] High-throughput sequencing technologies are intended to lower the cost of DNA sequencing beyond what is possible with standard dye-terminator methods. [79] In ultra-high-throughput sequencing as many as 500,000 sequencing-by-synthesis operations may be run in parallel.
First, DNA sequencing libraries are generated by clonal amplification by PCR in vitro. Second, the DNA is sequenced by synthesis, such that the DNA sequence is determined by the addition of nucleotides to the complementary strand rather than through chain-termination chemistry. Third, the spatially segregated, amplified DNA templates are ...
Dye-primer sequencing facilitates reading in an optical system for faster and more economical analysis and automation. The later development by Leroy Hood and coworkers [7] [8] of fluorescently labeled ddNTPs and primers set the stage for automated, high-throughput DNA sequencing. Sequence ladder by radioactive sequencing compared to ...
The column format is best suited for research and large scale applications where a high-throughput is not required. [102] Multi-well plate format is designed specifically for high-throughput synthesis on small scale to satisfy the growing demand of industry and academia for synthetic oligonucleotides. [103]
The ideal size of DNA fragments for the sequencing library depends on the sequencing platform that will be used. [4] [16] DNA can first be sheared to fragments around 300–500 bp long using sonication. [4] [16] [17] Fragments of this size are suitable for high-throughput sequencing.
Because artificial gene synthesis does not require template DNA, it is theoretically possible to make a completely synthetic DNA molecule with no limits on the nucleotide sequence or size. Synthesis of the first complete gene, a yeast tRNA , was demonstrated by Har Gobind Khorana and coworkers in 1972. [ 2 ]
The technology leading to these DNA sequencers was first released by Solexa in 2006 as the Genome Analyzer. [10] Illumina purchased Solexa in 2007. The Genome Analyzer uses a sequencing by synthesis method. The first model produced 1G per run. During the year 2009 the output was increased from 20G per run in August to 50G per run in December.
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