Sequencing-by-synthesis

Mechanism Dideoxy chain termination Pyrosequencing Sequencing by synthesis (SBS) Single molecule real-time SBS Semiconductor-based SBS... 

A heterogeneous assay was developed by Kartalov and Quake for DNA sequencing-by-synthesis to reliably sequence up to four consecutive base pairs [374], Briefly, the... 

Sequencing by synthesis Templeted enzymatic incorpora- tion F attached to bases of nucleoside triphosphates Fluorescence sensing, multicolour detection 43... 

The NGS techniques relying on sequencing-by-synthesis require the use of a DNA polymerase during the sequencing run. Sequencing-by-synthesis can be divided into two major techniques single-nucleotide addition and cyclic-reversible termination. 

The Illumina sequencing platform, which currently is the most widely used NGS platform, was commercialized in 2006, with Illumina acquiring Solexa in early 2007 (34). It is based on sequencing-by-synthesis chemistry, with reversible terminator nucleotides for the four bases, each labeled with a different fluorescent dye, and a DNA polymerase enzyme to incorporate them (35). This reaction mixture for the sequencing reactions is supplied onto the polonies created on a surface by solid-phase amplification. After each terminator nucleotide is incorporated into the DNA strand, a fluorescent signal is... 

Pyrosequencing is based on sequencing by synthesis. The assay takes advantage of the natural release of pyrophosphate whenever a nucleotide is incorporated onto an open 3 DNA strand. The released p5u-ophosphate is used in a sulfurylase reaction releasing ATP. The released ATP can be used by luciferase in the conversion of luciferin to oxyluciferin. The reaction results in the emission of light, which is collected by a CCD camera and recorded in the form of peaks, known as pyrograms (Fig. 1). When a nucleotide is not incorporated... 

Seo, T. S. et al. Four-color DNA sequencing by synthesis on a chip using photocleavable fluorescent nucleotides. Proceedings of the National Academy of Sciences of the United States of America 102, 5926-5931 (2005). 

Base-modified pyrophosphorylated nucleosides. A major contribution to advances in DNA sequencing has been achieved by a team of researchers from Columbia University. They developed a method which combined the Sanger dideoxynucleotide terminating reaction process to that of DNA sequencing by synthesis on a solid surface during polymerase chain reaction. The latter approach allows for deciphering many DNA... 

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