Polymerase Chain Reaction
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Polymerase Chain Reaction
The polymerase chain reaction (PCR) is a method widely used to make millions to billions of copies of a specific DNA sample rapidly, allowing scientists to amplify a very small sample of DNA (or a part of it) sufficiently to enable detailed study. PCR was invented in 1983 by American biochemist Kary Mullis at Cetus Corporation. Mullis and biochemist Michael Smith (chemist), Michael Smith, who had developed other essential ways of manipulating DNA, were jointly awarded the Nobel Prize in Chemistry in 1993. PCR is fundamental to many of the procedures used in genetic testing and research, including analysis of Ancient DNA, ancient samples of DNA and identification of infectious agents. Using PCR, copies of very small amounts of DNA sequences are exponentially amplified in a series of cycles of temperature changes. PCR is now a common and often indispensable technique used in medical laboratory research for a broad variety of applications including biomedical research and forensic ...
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PCR Tubes
PCR or pcr may refer to: Science and medicine * Pathologic complete response (pCR), in neoadjuvant therapy * Polymerase chain reaction ** COVID-19 testing, often performed using the polymerase chain reaction method * Phosphocreatine, a phosphorylated creatine molecule * Principal component regression, a statistical technique * Protein/creatinine ratio, in urine Technology * Passport Carrier Release, telecommunications software * Peak cell rate, on ATM networks * Platform Configuration Register, a Trusted Execution Technology implemented using a TPM * Processor Control Region, a Windows data structure * Program clock reference, in MPEG transport streams * XM PCR, a satellite receiver Political parties * ''Parti Communiste Réunionnais'' or Communist Party of Réunion * ''Partido Comunista Revolucionário'' or Revolutionary Communist Party * ''Partido Cívico Renovador'' or Civic Renovation Party, Dominican Republic * ''Partidul Comunist Român'' or Romanian Communist Party Other u ...
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Primer (molecular Biology)
A primer is a short, single-stranded nucleic acid used by all living organisms in the initiation of DNA synthesis. A synthetic primer is a type of oligo, short for oligonucleotide. DNA polymerases (responsible for DNA replication) are only capable of adding nucleotides to the 3’-end of an existing nucleic acid, requiring a primer be bound to the template before DNA polymerase can begin a complementary strand. DNA polymerase adds nucleotides after binding to the RNA primer and synthesizes the whole strand. Later, the RNA strands must be removed accurately and replaced with DNA nucleotides. This forms a gap region known as a nick that is filled in using a ligase. The removal process of the RNA primer requires several enzymes, such as Fen1, Lig1, and others that work in coordination with DNA polymerase, to ensure the removal of the RNA nucleotides and the addition of DNA nucleotides. Living organisms use solely RNA primers, while laboratory techniques in biochemistry and mole ...
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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, thymine, cytosine, and guanine. 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, Genographic Project, 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 advancements in DNA seque ...
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DNA Cloning
Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. The use of the word ''cloning'' refers to the fact that the method involves the replication of one molecule to produce a population of cells with identical DNA molecules. Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA. Molecular cloning methods are central to many contemporary areas of modern biology and medicine. In a conventional molecular cloning experiment, the DNA to be cloned is obtained from an organism of interest, then treated with enzymes in the test tube to generate smaller DNA fragments. Subsequently, these fragments are then combined with vector DNA to generate recombinant DNA molecules. The recombinant DNA is then ...
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Thermus Aquaticus
''Thermus aquaticus'' is a species of bacteria that can tolerate high temperatures, one of several thermophile, thermophilic bacteria that belong to the ''Deinococcota'' phylum. It is the source of the heat-resistant enzyme Taq polymerase, ''Taq'' DNA polymerase, one of the most important enzymes in molecular biology because of its use in the polymerase chain reaction (PCR) DNA amplification technique. History When studies of biological organisms in hot springs began in the 1960s, scientists thought that the life of thermophilic bacteria could not be sustained in temperatures above about . Soon, however, it was discovered that many bacteria in different springs not only survived, but also thrived in higher temperatures. In 1969, Thomas D. Brock and Hudson Freeze of Indiana University Bloomington, Indiana University reported a new species of thermophile, thermophilic bacteria which they named ''Thermus aquaticus''. The bacterium was first isolated from Mushroom Spring in the Geot ...
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Thermophile
A thermophile is a type of extremophile that thrives at relatively high temperatures, between . Many thermophiles are archaea, though some of them are bacteria and fungi. Thermophilic eubacteria are suggested to have been among the earliest bacteria. Thermophiles are found in geothermally heated regions of the Earth, such as hot springs like those in Yellowstone National Park and deep sea hydrothermal vents, as well as decaying plant matter, such as peat bogs and compost. They can survive at high temperatures, whereas other bacteria or archaea would be damaged and sometimes killed if exposed to the same temperatures. The enzymes in thermophiles function at high temperatures. Some of these enzymes are used in molecular biology, for example the ''Taq'' polymerase used in PCR. "Thermophile" is derived from the (''thermotita''), meaning heat, and (''philia''), love. Comparative surveys suggest that thermophile diversity is principally driven by pH, not temperature. Classificat ...
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Taq Polymerase
''Taq'' polymerase is a thermostable DNA polymerase I named after the thermophilic eubacterial microorganism ''Thermus aquaticus,'' from which it was originally isolated by master's student Alice Chien et al. in 1976. Its name is often abbreviated to ''Taq'' or ''Taq'' pol. It is frequently used in the polymerase chain reaction (PCR), a method for greatly amplifying the quantity of short segments of DNA. ''T. aquaticus'' is a bacterium that lives in hot springs and hydrothermal vents, and ''Taq'' polymerase was identified as an enzyme able to withstand the protein-denaturing conditions (high temperature) required during PCR. Therefore, it replaced the DNA polymerase from '' E. coli'' originally used in PCR. Enzymatic properties ''Taqs optimum temperature for activity is 75–80 °C, with a half-life of greater than 2 hours at 92.5 °C, 40 minutes at 95 °C and 9 minutes at 97.5 °C, and can replicate a 1000 base pair strand of DNA in less than 10 ...
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Thermostable DNA Polymerase
Thermostable DNA polymerases are DNA polymerases that originate from thermophiles, usually bacterial or archaeal species, and are therefore thermostable. They are used for the polymerase chain reaction and related methods for the amplification and modification of DNA. Properties Several DNA polymerases have been described with distinct properties that define their specific utilisation in a PCR, in real-time PCR or in an isothermal amplification. Being DNA polymerases, the thermostable DNA polymerases all have a 5'→3' polymerase activity, and either a 5'→3' or a 3'→5' exonuclease activity. Structure DNA polymerases are roughly shaped like a hand with a thumb, palm and fingers.T. A. Steitz''DNA polymerases: structural diversity and common mechanisms.''In: '' Journal of Biological Chemistry.'' Volume 274, issue 25, June 1999, p. 17395–17398, , .P. J. Rothwell, G. Waksman: ''Structure and mechanism of DNA polymerases.'' In: ''Advances in protein chemistry.'' Volu ...
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Exponential Growth
Exponential growth occurs when a quantity grows as an exponential function of time. The quantity grows at a rate directly proportional to its present size. For example, when it is 3 times as big as it is now, it will be growing 3 times as fast as it is now. In more technical language, its instantaneous rate of change (that is, the derivative) of a quantity with respect to an independent variable is proportional to the quantity itself. Often the independent variable is time. Described as a function, a quantity undergoing exponential growth is an exponential function of time, that is, the variable representing time is the exponent (in contrast to other types of growth, such as quadratic growth). Exponential growth is the inverse of logarithmic growth. Not all cases of growth at an always increasing rate are instances of exponential growth. For example the function f(x) = x^3 grows at an ever increasing rate, but is much slower than growing exponentially. For example, w ...
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Chain Reaction
A chain reaction is a sequence of reactions where a reactive product or by-product causes additional reactions to take place. In a chain reaction, positive feedback leads to a self-amplifying chain of events. Chain reactions are one way that systems which are not in thermodynamic equilibrium can release energy or increase entropy in order to reach a state of higher entropy. For example, a system may not be able to reach a lower energy state by releasing energy into the environment, because it is hindered or prevented in some way from taking the path that will result in the energy release. If a reaction results in a small energy release making way for more energy releases in an expanding chain, then the system will typically collapse explosively until much or all of the stored energy has been released. A macroscopic metaphor for chain reactions is thus a snowball causing a larger snowball until finally an avalanche results (" snowball effect"). This is a result of stored gravitati ...
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Nucleotide
Nucleotides are Organic compound, organic molecules composed of a nitrogenous base, a pentose sugar and a phosphate. They serve as monomeric units of the nucleic acid polymers – deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), both of which are essential biomolecules within all Life, life-forms on Earth. Nucleotides are obtained in the diet and are also synthesized from common Nutrient, nutrients by the liver. Nucleotides are composed of three subunit molecules: a nucleobase, a pentose, five-carbon sugar (ribose or deoxyribose), and a phosphate group consisting of one to three phosphates. The four nucleobases in DNA are guanine, adenine, cytosine, and thymine; in RNA, uracil is used in place of thymine. Nucleotides also play a central role in metabolism at a fundamental, cellular level. They provide chemical energy—in the form of the nucleoside triphosphates, adenosine triphosphate (ATP), guanosine triphosphate (GTP), cytidine triphosphate (CTP), and uridine triph ...
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Denaturation (biochemistry)
In biochemistry, denaturation is a process in which proteins or nucleic acids lose folded structure present in their native state due to various factors, including 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, 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 or dissociation of cofactors to aggregation due to the exposure of hydrophobic groups. The loss of solubility as a result of denaturation is called ''coagulation''. Denatured proteins, e.g., metalloenzymes, lose their 3D structure or metal cofactor, and therefore, cannot function. Proper protein folding is key to whether a globular or memb ...
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