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Polymerase chain reaction functions as an in vitro DNA cloning technique
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Peer-reviewed literature and reference materials establish that the polymerase chain reaction (PCR) functions as a method for amplifying DNA in vitro, serving as an in vitro cloning technique.

Evidence for · 6
1990 · cited by 292
In the polymerase chain reaction (PCR) technique, DNA is amplified in vitro by a series of polymerization cycles consisting of three temperature-dependent steps: DNA denaturation, primer-template annealing, and DNA synthesis by a thermostable DNA polymerase. The purity and yield of the reaction products depend on several parameters, one of which is the annealing temperature (Ta). At both sub- and super-optimal Ta values, non-specific products may be formed, and the yield of products is reduced. Optimizing the Ta is especially critical when long products are synthesized or when total genomic DNA is the substrate for PCR. In this article we experimentally determine the optimal annealing temperature (TaOPT) values for several primer-template pairs and develop a method for its calculation. The TaOPT is found to be a function of the melting temperatures of the less stable primer-template pair and of the product. The fact that experimental and calculated TaOPT values agree to within 0.7 degree C eliminates the need for determining TaOPT experimentally. Synthesis of DNA fragments shorter than 1 kb is more efficient if a variable Ta is used, such that the Ta is higher in each consecutive cycle.
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The polymerase chain reaction. The polymerase chain reaction (PCR) is a powerful new method for 'in vitro cloning'. It can selectively amplify a single molecule of template DNA several millionfold in a few hours and has made possible new approaches to problems in molecular genetics, evolutionary biology, and development. Published in Trends in genetics : TIG (1989)
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commonly replicate DNA in vitro using the polymerase chain reaction (PCR). PCR uses a pair of primers to span a target region in template DNA, and then polymerizes DNA replication is the process by which a cell makes exact copies of its DNA. This process occurs in all organisms and is essential to biological inheritance, cell division, and repair of damaged tissues. DNA replication ensures that each of the newly divided daughter cells receives its own copy of each DNA molecule. DNA most commonly occurs in double-stranded form, made up of two complementary st DNA replication is the process by which a cell makes exact copies of its DNA. This process occurs in all organisms and is essential to biological inheritance, cell division, and repair of damaged tissues. DNA replication ensures that each of the newly divided daughter cells receives its own copy of each DNA molecule. DNA most commonly occurs in double-stranded form, made up of two complementary strands held together by base pairing of the nucleotides comprising each strand. The two linear strands of a double-stranded DNA molecule typically twist together in the shape of a double helix. During replication, the two strands are separated, and each strand of the original DNA molecule then serves as a template for the production of a complementary counterpart strand, a process referred to as semiconservative replication. As a result, each replicated DNA molecule is composed of one original DNA strand as well as one newly synthesized strand. Cellular proofreading and error-checking mechanisms ensure near-perfect fidelity for DNA replication. DNA replication usually begins at specific locations known as origins of replication which are scattered across the genome. Unwinding of DNA at the origin is accommodated by enzymes known as helicases and results in replication forks growing bi-directionally from the origin. Numerous proteins are associated with the replication fork to help in the initiation and continuation of DNA synthesis. Most prominently, DNA polymerase synthesizes the new strands by incorporating nucleotides that complement the nucleotides of the template strand. DNA replication occurs during the S (synthesis) stage of interphase. DNA replication can also be performed in vitro (artificially, outside a cell). DNA polymerases isolated from cells and artificial DNA primers can be used to start DNA synthesis at known sequences in a template DNA molecule. Polymerase chain reaction (PCR), ligase chain reaction (LCR), and transcription-mediated amplification (TMA) are all common examples of this technique. In March 2021, researchers reported evidence suggesting that a preliminary form of transfer RNA, a necessary component of translation (the biological synthesis…
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La reacción en cadena de la polimerasa o PCR (siglas de su nombre en inglés Polymerase Chain Reaction) permite generar una gran cantidad de copias de un fragmento de DNA (ácido desoxirribonulceico). El requisito fundamental para poder llevar a cabo la reacción es disponer de fragmentos cortos de DNA de cadena sencilla complementarios a los extremos del fragmento a amplificar. Estos fragmentos servirán como cebadores para que una enzima polimerasa sea capaz de incorporar nucleótidos complementarios a la cadena molde. Una vez completada la reacción la cantidad fragmento amplificado se puede visu
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Production of discrete high specific activity DNA probes using the polymerase chain reaction. Conditions are described for the synthesis of discrete DNA probes with high specific activity using the polymerase chain reaction. This method enables the production of DNA probes between any two oligonucleotide sequences from cloned or uncloned templates. These probes are uniform in length and their specific activity (1 x 10(9) cpm/microgram) is comparable to probes produced by other methods. Published in Gene analysis techniques
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Use of the DNA polymerase chain reaction for homology probing: isolation of partial cDNA or genomic clones encoding the iron-sulfur protein of succinate dehydrogenase from several species. The DNA polymerase chain reaction was developed for in vitro amplification of specific DNA sequences, and it has been used for a wide variety of purposes in several fields. We have developed an application of the polymerase chain reaction that is useful for the isolation of partial cDNA or genomic clones of conserved genes. We used this technique to clone the gene encoding the iron protein subunit (27 kDa) of succinate dehydrogenase (EC 1.3.5.1) from several species, including human, rat, Drosophila melanogaster, Arabidopsis thaliana, Schizosaccharomyces pombe, and Saccharomyces cerevisiae. Mixed oligonucleotide primers corresponding to two conserved regions of the protein were used in conjunction with genomic and cDNA templates in the reaction. The primers contained all possible nucleotide combinations that could encode the corresponding peptide sequences. These oligonucleotide mixtures contained 262,144 (2(18] and 8192 (2(13] unique sequences, respectively.
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