Second-generation Sequencing
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Second-generation Sequencing
Massive parallel sequencing or massively parallel sequencing is any of several high-throughput approaches to DNA sequencing using the concept of massively parallel processing; it is also called next-generation sequencing (NGS) or second-generation sequencing. Some of these technologies emerged between 1994 and 1998 and have been commercially available since 2005. These technologies use miniaturized and parallelized platforms for sequencing of 1 million to 43 billion short reads (50 to 400 bases each) per instrument run. Many NGS platforms differ in engineering configurations and sequencing chemistry. They share the technical paradigm of massive parallel sequencing via spatially separated, clonally amplified DNA templates or single DNA molecules in a flow cell. This design is very different from that of Sanger sequencing—also known as capillary sequencing or first-generation sequencing—which is based on electrophoretic separation of chain-termination products produced in individ ...
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DNA Sequencing
DNA sequencing is the process of determining the nucleic acid sequence – the order of nucleotides in DNA. It includes any method or technology that is used to determine the order of the four bases: adenine, guanine, cytosine, and thymine. The advent of rapid DNA sequencing methods has greatly accelerated biological and medical research and discovery. Knowledge of DNA sequences has become indispensable for basic biological research, DNA Genographic Projects and in numerous applied fields such as medical diagnosis, biotechnology, forensic biology, virology and biological systematics. Comparing healthy and mutated DNA sequences can diagnose different diseases including various cancers, characterize antibody repertoire, and can be used to guide patient treatment. Having a quick way to sequence DNA allows for faster and more individualized medical care to be administered, and for more organisms to be identified and cataloged. The rapid speed of sequencing attained with modern D ...
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Oligonucleotide
Oligonucleotides are short DNA or RNA molecules, oligomers, that have a wide range of applications in genetic testing, research, and forensics. Commonly made in the laboratory by solid-phase chemical synthesis, these small bits of nucleic acids can be manufactured as single-stranded molecules with any user-specified sequence, and so are vital for artificial gene synthesis, polymerase chain reaction (PCR), DNA sequencing, molecular cloning and as molecular probes. In nature, oligonucleotides are usually found as small RNA molecules that function in the regulation of gene expression (e.g. microRNA), or are degradation intermediates derived from the breakdown of larger nucleic acid molecules. Oligonucleotides are characterized by the sequence of nucleotide residues that make up the entire molecule. The length of the oligonucleotide is usually denoted by " -mer" (from Greek ''meros'', "part"). For example, an oligonucleotide of six nucleotides (nt) is a hexamer, while one of 25 nt wou ...
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Pyrosequencing
Pyrosequencing is a method of DNA sequencing (determining the order of nucleotides in DNA) based on the "sequencing by synthesis" principle, in which the sequencing is performed by detecting the nucleotide incorporated by a DNA polymerase. Pyrosequencing relies on light detection based on a chain reaction when pyrophosphate is released. Hence, the name pyrosequencing. The principle of pyrosequencing was first described in 1993 by, Bertil Pettersson, Mathias Uhlen and Pål Nyren by combining the solid phase sequencing method using streptavidin coated magnetic beads with recombinant DNA polymerase lacking 3´to 5´exonuclease activity (proof-reading) and luminescence detection using the firefly luciferase enzyme. A mixture of three enzymes (DNA polymerase, ATP sulfurylase and firefly luciferase) and a nucleotide (dNTP) are added to single stranded DNA to be sequenced and the incorporation of nucleotide is followed by measuring the light emitted. The intensity of the light determi ...
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Stockholm
Stockholm () is the Capital city, capital and List of urban areas in Sweden by population, largest city of Sweden as well as the List of urban areas in the Nordic countries, largest urban area in Scandinavia. Approximately 980,000 people live in the Stockholm Municipality, municipality, with 1.6 million in the Stockholm urban area, urban area, and 2.4 million in the Metropolitan Stockholm, metropolitan area. The city stretches across fourteen islands where Mälaren, Lake Mälaren flows into the Baltic Sea. Outside the city to the east, and along the coast, is the island chain of the Stockholm archipelago. The area has been settled since the Stone Age, in the 6th millennium BC, and was founded as a city in 1252 by Swedish statesman Birger Jarl. It is also the county seat of Stockholm County. For several hundred years, Stockholm was the capital of Finland as well (), which then was a part of Sweden. The population of the municipality of Stockholm is expected to reach o ...
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Mostafa Ronaghi
Mostafa Ronaghi ( fa, مصطفی رونقی; born 1968) is an Iranian molecular biologist, specializing in DNA sequencing methodology. He earned his Ph.D. from the Royal Institute of Technology in Sweden in 1998. , he is the Chief Technology Officer and Senior Vice President at Illumina. Prior to this position, he was a principal investigator and Senior Research Associate at the Stanford Genome Technology Center at Stanford University, focusing on developing analytical techniques for molecular diagnostics. He is principal investigator for several grants including grants from the National Human Genome Research Institute (NHGRI), part of the National Institutes of Health for the development of array-based Pyrosequencing. In 1998, he described together with Pål Nyren anMathias Uhlena solution-based variant of the pyrosequencing Pyrosequencing is a method of DNA sequencing (determining the order of nucleotides in DNA) based on the "sequencing by synthesis" principle, in which t ...
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Pål Nyrén
Pål Nyrén (born 1955) is a biochemistry professor at the Royal Institute of Technology (KTH), Stockholm. He is most famous for developing the pyrosequencing method for DNA sequencing. Career *1999 Professor in Biochemistry, KTH, Stockholm *1997 Founder of the company Biotage AB (former Pyrosequencing AB) *1988 Associate professor (Docent) Biochemistry, University of Stockholm *1985-86 Postdoc at LMB, MRC, Cambridge, G.B. with prof John Walker *1985 PhD (Tekn. Doktor) Biochemistry, University of Stockholm (Thesis title: "The proton pumping pyrophosphatase from ''Rhodospirillum rubrum''") *1981 MSc (Civ. ing.) Chemical Engineering, KTH, Stockholm Recognition *2013 winner of the European Inventor Award in the SMEs categoryInvention


Whitehead Institute For Biomedical Research
Whitehead Institute for Biomedical Research is a non-profit research institute located in Cambridge, Massachusetts, United States that is dedicated to improving human health through basic biomedical research. It was founded as a fiscally independent entity from the Massachusetts Institute of Technology (MIT), where its 18 members all hold faculty appointments in the MIT Department of Biology or the MIT Department of Bioengineering. Two members (Rudolf Jaenisch, 2010, and Robert Weinberg, 1997) are National Medal of Science recipients; ten have been elected to the National Academy of Sciences; and four have been elected to the National Academy of Medicine; six are Howard Hughes Medical Institute Investigators. In September 2019, Ruth Lehmann was announced as the new director of the Whitehead Institute for Biomedical Research, commencing July 2020 and succeeding David Page. History Whitehead Institute was founded in 1982 by industrialist and philanthropist Edwin C. “Jack ...
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Christopher Adams (scientist)
Christopher P. Adams is an American scientist, entrepreneur, and inventor who founded and led (as chief executive officer) multiple biotechnology companies, including Mosaic Technologies and Andarix Pharmaceuticals. Adams has made a notable contribution to the field of genetics as a co-inventor (with Steve Kron) of " bridge amplification," a polymerase chain reaction (PCR) technique that paved the way for development of DNA sequencing and genome sequencing. Geneticist George Church and computational biologist Rob Mitra adapted Adams' and Kron's technique to be used for clonal amplification. Adams has spoken about the difficulties he faced in launching Mosaic and getting investors to buy in. He cites being "an African American without a graduate degree" as a major reason he faced skepticism and rejection while attempting to start Mosaic, and that his persistence was a key factor that helped him "overcome investors' reluctance." Adams has said:"Keep in mind, I had no Ph.D. and no MBA ...
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Pascal Mayer
Pascal Mayer is a french scientist who was awarded Breakthrough Prize in Life Sciences in 2022 for researching paved way for inexpensive and rapid DNA sequencing DNA sequencing is the process of determining the nucleic acid sequence – the order of nucleotides in DNA. It includes any method or technology that is used to determine the order of the four bases: adenine, guanine, cytosine, and thymine. Th ... around the world. References {{Authority control French scientists Living people 1963 births ...
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Denaturation (biochemistry)
In biochemistry, denaturation is a process in which proteins or nucleic acids lose the quaternary structure, tertiary structure, and secondary structure which is present in their native state, by application of some external stress or compound such as a strong acid or base, a concentrated inorganic salt, an organic solvent (e.g., alcohol or chloroform), agitation and radiation or heat. If proteins in a living cell are denatured, this results in disruption of cell activity and possibly cell death. Protein denaturation is also a consequence of cell death. Denatured proteins can exhibit a wide range of characteristics, from conformational change and loss of solubility to aggregation due to the exposure of hydrophobic groups. The loss of solubility as a result of denaturation is called ''coagulation''. Denatured proteins lose their 3D structure and therefore cannot function. Protein folding is key to whether a globular or membrane protein can do its job correctly; it must be ...
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Oligonucleotide
Oligonucleotides are short DNA or RNA molecules, oligomers, that have a wide range of applications in genetic testing, research, and forensics. Commonly made in the laboratory by solid-phase chemical synthesis, these small bits of nucleic acids can be manufactured as single-stranded molecules with any user-specified sequence, and so are vital for artificial gene synthesis, polymerase chain reaction (PCR), DNA sequencing, molecular cloning and as molecular probes. In nature, oligonucleotides are usually found as small RNA molecules that function in the regulation of gene expression (e.g. microRNA), or are degradation intermediates derived from the breakdown of larger nucleic acid molecules. Oligonucleotides are characterized by the sequence of nucleotide residues that make up the entire molecule. The length of the oligonucleotide is usually denoted by " -mer" (from Greek ''meros'', "part"). For example, an oligonucleotide of six nucleotides (nt) is a hexamer, while one of 25 nt wou ...
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Polymerase
A polymerase is an enzyme ( EC 2.7.7.6/7/19/48/49) that synthesizes long chains of polymers or nucleic acids. DNA polymerase and RNA polymerase are used to assemble DNA and RNA molecules, respectively, by copying a DNA template strand using base-pairing interactions or RNA by half ladder replication. A DNA polymerase from the thermophilic bacterium, ''Thermus aquaticus'' (''Taq'') ( PDBbr>1BGX EC 2.7.7.7) is used in the polymerase chain reaction, an important technique of molecular biology. A polymerase may be template dependent or template independent. Poly-A-polymerase is an example of template independent polymerase. Terminal deoxynucleotidyl transferase also known to have template independent and template dependent activities. Types By function *DNA polymerase (DNA-directed DNA polymerase, DdDP) **Family A: DNA polymerase I; Pol γ, θ, ν **Family B: DNA polymerase II; Pol α, δ, ε, ζ **Family C: DNA polymerase III holoenzyme **Family X: Pol β, λ, μ * ...
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