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Illumina dye sequencing
Illumina dye sequencing is a technique used to determine the series of base pairs in DNA, also known as DNA sequencing. It was based on inventions of S Balasubramanian and D Klenerman of Cambridge University,〔http://technology.illumina.com/technology/next-generation-sequencing/solexa-technology.html〕 who subsequently founded Solexa, a company later acquired by Illumina. This sequencing method is based on reversible dye-terminators that enable the identification of single bases as they are introduced into DNA strands. It is often employed to sequence difficult regions, such as homopolymers and repetitive sequences. It can also be used for whole-genome and region sequencing, transcriptome analysis, metagenomics, small RNA discovery, methylation profiling, and genome-wide protein-nucleic acid interaction analysis.〔(- Sequencing Technology )〕〔Meyer M., Kircher M.(2010).Illumina Sequencing Library Preparation for Highly Multiplexed Target Capture and Sequencing. ''Cold Springs Harbor Protocols".doi:10.1101/pdb.prot5448.〕
==Overview==
Before sequencing, DNA must be purified. The next step after purification is tagmentation. Enzymes called transposomes randomly cut the DNA into short segments (“tags”). Adapters are added on either side of the cut points (ligation). Strands that fail to have adapters ligated are washed away.〔(【引用サイトリンク】url= )〕
Illumina dye sequencing begins with the attachment of DNA molecules to primers on a slide, followed by amplification of that DNA to produce local colonies. This "DNA cluster" generation technology had been invented and developed in late 1996 at Glaxo Wellcome's Geneva Biomedical Research Institute (GBRI), by Dr Pascal Mayer and Dr Laurent Farinelli,〔patents WO 9844151, WO 9844152〕 and was publicly presented for the first time in 1998.〔(DNA colony massively parallel sequencing ams98 presentation )〕 Solexa's proprietary chemistry, which also required an engineered polymerase, was developed at its site in Gt. Chesterford near Cambridge (UK), the initial commercial versions of the clusters technology were also perfected there. The four types (adenine, cytosine, guanine, and thymine) of reversible terminate bases are added, each fluorescently labeled with a different color and attached with a blocking group. The four bases then compete for binding sites on the template DNA to be sequenced and non-incorporated molecules are washed away. After each synthesis, a laser is used to excite the dyes and a photograph of the incorporated base is taken. A chemical deblocking step is then used in the removal of the 3’ terminal blocking group and the dye in a single step. The process is repeated until the full DNA molecule is sequenced.〔
抄文引用元・出典: フリー百科事典『 ウィキペディア（Wikipedia）』
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