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Polymerase Cycling Assembly
Polymerase cycling assembly (or PCA, also known as Assembly PCR) is a method for the assembly of large DNA oligonucleotides from shorter fragments. The process uses the same technology as PCR, but takes advantage of DNA hybridization and annealing as well as DNA polymerase to amplify a complete sequence of DNA in a precise order based on the single stranded oligonucleotides used in the process. It thus allows for the production of synthetic genes and even entire synthetic genomes. PCA principles Much like how primers are designed such that there is a forward primer and a reverse primer capable of allowing DNA polymerase to fill the entire template sequence, PCA uses the same technology but with multiple oligonucleotides. While in PCR the customary size of oligonucleotides used is 18 base pairs, in PCA lengths of up to 50 are used to ensure uniqueness and correct hybridization. Each oligonucleotide is designed to be either part of the top or bottom strand of the target sequence ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Oligonucleotide
Oligonucleotides are short DNA or RNA molecules, oligomers, that have a wide range of applications in genetic testing, Recombinant DNA, research, and Forensic DNA, forensics. Commonly made in the laboratory by Oligonucleotide synthesis, solid-phase chemical synthesis, these small fragments 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 Fluorescence in situ hybridization, 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 Nucleic acid sequence, sequence of nucleotide residues that make up the entire molecule. The length of the oligonucleotide is usually denoted by "Monomer, -m ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
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 ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
DNA Polymerase
A DNA polymerase is a member of a family of enzymes that catalyze the synthesis of DNA molecules from nucleoside triphosphates, the molecular precursors of DNA. These enzymes are essential for DNA replication and usually work in groups to create two identical DNA duplexes from a single original DNA duplex. During this process, DNA polymerase "reads" the existing DNA strands to create two new strands that match the existing ones. These enzymes catalysis, catalyze the chemical reaction : deoxynucleoside triphosphate + DNAn pyrophosphate + DNAn+1. DNA polymerase adds nucleotides to the Directionality (molecular biology), three prime (3')-end of a DNA strand, one nucleotide at a time. Every time a Cell division, cell divides, DNA polymerases are required to duplicate the cell's DNA, so that a copy of the original DNA molecule can be passed to each daughter cell. In this way, genetic information is passed down from generation to generation. Before replication can take place, an enzy ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Synthetic Gene
Artificial gene synthesis, or simply gene synthesis, refers to a group of methods that are used in synthetic biology to construct and assemble genes from nucleotides ''de novo''. Unlike DNA synthesis in living cells, artificial gene synthesis does not require template DNA, allowing virtually any DNA sequence to be synthesized in the laboratory. It comprises two main steps, the first of which is solid-phase DNA synthesis, sometimes known as DNA printing. This produces oligonucleotide fragments that are generally under 200 base pairs. The second step then involves connecting these oligonucleotide fragments using various DNA assembly methods. Because artificial gene synthesis does not require template DNA, it is theoretically possible to make a completely synthetic DNA molecule with no limits on the nucleotide sequence or size. Synthesis of the first complete gene, a yeast tRNA, was demonstrated by Har Gobind Khorana and coworkers in 1972. Synthesis of the first peptide- and protein-c ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Genome
A genome is all the genetic information of an organism. It consists of nucleotide sequences of DNA (or RNA in RNA viruses). The nuclear genome includes protein-coding genes and non-coding genes, other functional regions of the genome such as regulatory sequences (see non-coding DNA), and often a substantial fraction of junk DNA with no evident function. Almost all eukaryotes have mitochondrial DNA, mitochondria and a small mitochondrial genome. Algae and plants also contain chloroplast DNA, chloroplasts with a chloroplast genome. The study of the genome is called genomics. The genomes of many organisms have been Whole-genome sequencing, sequenced and various regions have been annotated. The first genome to be sequenced was that of the virus φX174 in 1977; the first genome sequence of a prokaryote (''Haemophilus influenzae'') was published in 1995; the yeast (''Saccharomyces cerevisiae'') genome was the first eukaryotic genome to be sequenced in 1996. The Human Genome Project ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
PCA Polymerase Cycling Assembly
PCA may refer to: Medicine and biology * Patient-controlled analgesia * Plate count agar in microbiology * Polymerase cycling assembly, for large DNA oligonucleotides * Posterior cerebral artery * Posterior cortical atrophy, a form of dementia *Prostate cancer * Protein-fragment complementation assay, to identify protein–protein interactions * Protocatechuic acid, a phenolic acid. * Personal Care Assistant, also known as unlicensed assistive personnel *Procainamide Military and government * EU-Armenia Partnership and Cooperation Agreement (PCA agreement between Armenia and the EU) * Parks Canada Agency * Partnership and Cooperation Agreement (EU) * Permanent change of assignment in US armed forces Organizations Business * Packaging Corporation of America * Peanut Corporation of America, former company * Pennsylvania Central Airlines 1936-1948 Education * Pacific Coast Academy, fictional school in TV series ''Zoey 101'' * Parents and citizens associations for ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Gibson Assembly
Gibson assembly is a molecular cloning method that allows for the joining of multiple DNA fragments in a single, isothermal reaction. It is named after its creator, Daniel G. Gibson, who is the chief technology officer and co-founder of the synthetic biology company Telesis Bio. The technology is more efficient than manual plasmid genetic recombination methods but remains expensive as it is still under patent. Process The entire Gibson assembly reaction requires few components with minor manipulations. The method can simultaneously combine up to 15 DNA fragments based on sequence identity. It requires that the DNA fragments contain ~20-40 base pair overlap with adjacent DNA fragments. These DNA fragments are mixed with a cocktail of three enzymes, along with other buffer components. The three required enzyme activities are: exonuclease, DNA polymerase, and DNA ligase. * The exonuclease chews back DNA from the 5' end, thus not inhibiting polymerase activity and allowing the re ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Kilobase
A base pair (bp) is a fundamental unit of double-stranded nucleic acids consisting of two nucleobases bound to each other by hydrogen bonds. They form the building blocks of the DNA double helix and contribute to the folded structure of both DNA and RNA. Dictated by specific hydrogen bonding patterns, "Watson–Crick" (or "Watson–Crick–Franklin") base pairs (guanine–cytosine and adenine– thymine) allow the DNA helix to maintain a regular helical structure that is subtly dependent on its nucleotide sequence. The complementary nature of this based-paired structure provides a redundant copy of the genetic information encoded within each strand of DNA. The regular structure and data redundancy provided by the DNA double helix make DNA well suited to the storage of genetic information, while base-pairing between DNA and incoming nucleotides provides the mechanism through which DNA polymerase replicates DNA and RNA polymerase transcribes DNA into RNA. Many DNA-binding protein ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Cambridge University
The University of Cambridge is a Public university, public collegiate university, collegiate research university in Cambridge, England. Founded in 1209, the University of Cambridge is the List of oldest universities in continuous operation, world's third-oldest university in continuous operation. The university's founding followed the arrival of scholars who left the University of Oxford for Cambridge after a dispute with local townspeople. The two ancient university, ancient English universities, although sometimes described as rivals, share many common features and are often jointly referred to as Oxbridge. In 1231, 22 years after its founding, the university was recognised with a royal charter, granted by Henry III of England, King Henry III. The University of Cambridge includes colleges of the University of Cambridge, 31 semi-autonomous constituent colleges and List of institutions of the University of Cambridge#Schools, Faculties, and Departments, over 150 academic departm ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
International Genetically Engineered Machine
The iGEM (International Genetically Engineered Machine) competition is a worldwide synthetic biology competition that was initially aimed at undergraduate and 'overgraduate' university students, but has since expanded to include divisions for high school students, entrepreneurs, and community laboratories. iGEM is presented as "''the'' ''heart of synthetic biology''" - educating the next generation of leaders and workforce of the field. Since its inception in 2003, over 80 000 students from over 65 countries have been trained in the responsible, safe and secure use of synthetic biology. The iGEM Competition is a flagship program of the iGEM Foundation - an independent, non-profit organization dedicated to the advancement of synthetic biology, education and competition, and the development of an open, collaborative, and cooperative community. Aside from the competition, iGEM has established many initiatives and programs to support the future growth of synthetic biology throughout t ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Amplifiers
An amplifier, electronic amplifier or (informally) amp is an electronic device that can increase the magnitude of a signal (a time-varying voltage or current). It is a two-port electronic circuit that uses electric power from a power supply to increase the amplitude (magnitude of the voltage or current) of a signal applied to its input terminals, producing a proportionally greater amplitude signal at its output. The amount of amplification provided by an amplifier is measured by its gain: the ratio of output voltage, current, or power to input. An amplifier is defined as a circuit that has a power gain greater than one. An amplifier can be either a separate piece of equipment or an electrical circuit contained within another device. Amplification is fundamental to modern electronics, and amplifiers are widely used in almost all electronic equipment. Amplifiers can be categorized in different ways. One is by the frequency of the electronic signal being amplified. For examp ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Genetic Engineering
Genetic engineering, also called genetic modification or genetic manipulation, is the modification and manipulation of an organism's genes using technology. It is a set of Genetic engineering techniques, technologies used to change the genetic makeup of cells, including the transfer of genes within and across species boundaries to produce improved or novel organisms. New DNA is obtained by either isolating and copying the genetic material of interest using recombinant DNA methods or by Artificial gene synthesis, artificially synthesising the DNA. A Vector (molecular biology), construct is usually created and used to insert this DNA into the host organism. The first recombinant DNA molecule was made by Paul Berg in 1972 by combining DNA from the monkey virus SV40 with the Lambda phage, lambda virus. As well as inserting genes, the process can be used to remove, or "Gene knockout, knock out", genes. The new DNA can either be inserted randomly or Gene targeting, targeted to a spe ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
