Reverse Transfection
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Reverse Transfection
Reverse transfection is a technique for the transfer of genetic material into cells. As DNA is printed on a glass slide for the transfection process (the deliberate introduction of nucleic acids into cells) to occur before the addition of adherent cells, the order of addition of DNA and adherent cells is reverse that of conventional transfection. Hence, the word “reverse” is used. Process Transfection-mix preparation for slide printing A DNA-gelatin mixture may be used for printing onto a slide. Gelatin powder is first dissolved in sterile Milli-Q water to form a 0.2% gelatin solution. Purified DNA plasmid is then mixed with the gelatin solution, and the final gelatin concentration is kept greater than 0.17%. Besides gelatin, atelocollagen and fibronectin are also successful transfection vectors for introducing foreign DNA into the cell nucleus. Slide printing of DNA-gelatin mixture After the DNA-gelatin mixture preparation, the mixture is pipetted onto a slide surfac ...
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Gene
In biology, the word gene (from , ; "...Wilhelm Johannsen coined the word gene to describe the Mendelian units of heredity..." meaning ''generation'' or ''birth'' or ''gender'') can have several different meanings. The Mendelian gene is a basic unit of heredity and the molecular gene is a sequence of nucleotides in DNA that is transcribed to produce a functional RNA. There are two types of molecular genes: protein-coding genes and noncoding genes. During gene expression, the DNA is first copied into RNA. The RNA can be directly functional or be the intermediate template for a protein that performs a function. The transmission of genes to an organism's offspring is the basis of the inheritance of phenotypic traits. These genes make up different DNA sequences called genotypes. Genotypes along with environmental and developmental factors determine what the phenotypes will be. Most biological traits are under the influence of polygenes (many different genes) as well as gen ...
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Incubator (culture)
An incubator is a device used to grow and maintain microbiological cultures or cell cultures. The incubator maintains optimal temperature, humidity and other conditions such as the CO2 and oxygen content of the atmosphere inside. Incubators are essential for much experimental work in cell biology, microbiology and molecular biology and are used to culture both bacterial and eukaryotic cells. An incubator is made up of a chamber with a regulated temperature. Some incubators also regulate humidity, gas composition, or ventilation within that chamber. The simplest incubators are insulated boxes with an adjustable heater, typically going up to 60 to 65 °C (140 to 150 °F), though some can go slightly higher (generally to no more than 100 °C). The most commonly used temperature both for bacteria such as the frequently used E. coli as well as for mammalian cells is approximately 37 °C (99 °F), as these organisms grow well under such conditions. For other organisms used in biologi ...
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Small Interfering RNA
Small interfering RNA (siRNA), sometimes known as short interfering RNA or silencing RNA, is a class of double-stranded RNA at first non-coding RNA molecules, typically 20-24 (normally 21) base pairs in length, similar to miRNA, and operating within the RNA interference (RNAi) pathway. It interferes with the expression of specific genes with complementary nucleotide sequences by degrading mRNA after transcription, preventing translation. Text was copied from this source, which is available under Creative Commons Attribution 4.0 International License Structure Naturally occurring siRNAs have a well-defined structure that is a short (usually 20 to 24- bp) double-stranded RNA (dsRNA) with phosphorylated 5' ends and hydroxylated 3' ends with two overhanging nucleotides. The Dicer enzyme catalyzes production of siRNAs from long dsRNAs and small hairpin RNAs. siRNAs can also be introduced into cells by transfection. Since in principle any gene can be knocked down by a syntheti ...
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Data Quality
Data quality refers to the state of qualitative or quantitative pieces of information. There are many definitions of data quality, but data is generally considered high quality if it is "fit for tsintended uses in operations, decision making and planning". Moreover, data is deemed of high quality if it correctly represents the real-world construct to which it refers. Furthermore, apart from these definitions, as the number of data sources increases, the question of internal data consistency becomes significant, regardless of fitness for use for any particular external purpose. People's views on data quality can often be in disagreement, even when discussing the same set of data used for the same purpose. When this is the case, data governance is used to form agreed upon definitions and standards for data quality. In such cases, data cleansing, including standardization, may be required in order to ensure data quality. Definitions Defining data quality is difficult due to the ma ...
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List Of Distinct Cell Types In The Adult Human Body
There are many different types of cells in the human body. Cells derived primarily from endoderm Exocrine secretory epithelial cells * Brunner's gland cell in duodenum (enzymes and alkaline mucus) *Insulated goblet cell of respiratory and digestive tracts (mucus secretion) *Stomach **Foveolar cell (mucus secretion) **Chief cell ( pepsinogen secretion) **Parietal cell (hydrochloric acid secretion) * Pancreatic acinar cell (bicarbonate and digestive enzyme secretion) *Paneth cell of small intestine (lysozyme secretion) *Type II pneumocyte of lung ( surfactant secretion) *Club cell of lung Barrier cells *Type I pneumocyte (lung) * Gall bladder epithelial cell * Centroacinar cell (pancreas) *Intercalated duct cell (pancreas) *Intestinal brush border cell (with microvilli) Hormone-secreting cells *Enteroendocrine cell **K cell (secretes gastric inhibitory peptide) **L cell (secretes glucagon-like peptide-1, peptide YY3-36, oxyntomodulin, and glucagon-like peptide-2) **I ...
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Time-lapse
Time-lapse photography is a technique in which the frequency at which film frames are captured (the frame rate) is much lower than the frequency used to view the sequence. When played at normal speed, time appears to be moving faster and thus ''lapsing''. For example, an image of a scene may be captured at 1 frame per second but then played back at 30 frames per second; the result is an apparent ''30 times'' speed increase. Similarly, film can also be played at a much lower rate than at which it was captured, which slows down an otherwise fast action, as in slow motion or high-speed photography. Processes that would normally appear subtle and slow to the human eye, such as the motion of the sun and stars in the sky or the growth of a plant, become very pronounced. Time-lapse is the extreme version of the cinematography technique of ''undercranking''. Stop motion animation is a comparable technique; a subject that does not actually move, such as a puppet, can repeatedly be move ...
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Gene Expression
Gene expression is the process by which information from a gene is used in the synthesis of a functional gene product that enables it to produce end products, protein or non-coding RNA, and ultimately affect a phenotype, as the final effect. These products are often proteins, but in non-protein-coding genes such as transfer RNA (tRNA) and small nuclear RNA (snRNA), the product is a functional non-coding RNA. Gene expression is summarized in the central dogma of molecular biology first formulated by Francis Crick in 1958, further developed in his 1970 article, and expanded by the subsequent discoveries of reverse transcription and RNA replication. The process of gene expression is used by all known life—eukaryotes (including multicellular organisms), prokaryotes (bacteria and archaea), and utilized by viruses—to generate the macromolecular machinery for life. In genetics, gene expression is the most fundamental level at which the genotype gives rise to the phenotype, '' ...
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High Content Screening
High-content screening (HCS), also known as high-content analysis (HCA) or cellomics, is a method that is used in biological research and drug discovery to identify substances such as small molecules, peptides, or RNAi that alter the phenotype of a cell in a desired manner. Hence high content screening is a type of phenotypic screen conducted in cells involving the analysis of whole cells or components of cells with simultaneous readout of several parameters. HCS is related to high-throughput screening (HTS), in which thousands of compounds are tested in parallel for their activity in one or more biological assays, but involves assays of more complex cellular phenotypes as outputs. Phenotypic changes may include increases or decreases in the production of cellular products such as proteins and/or changes in the morphology (visual appearance) of the cell. Hence HCA typically involves automated microscopy and image analysis. Unlike high-content analysis, high-content screening imp ...
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Eukaryote
Eukaryotes () are organisms whose cells have a nucleus. All animals, plants, fungi, and many unicellular organisms, are Eukaryotes. They belong to the group of organisms Eukaryota or Eukarya, which is one of the three domains of life. Bacteria and Archaea (both prokaryotes) make up the other two domains. The eukaryotes are usually now regarded as having emerged in the Archaea or as a sister of the Asgard archaea. This implies that there are only two domains of life, Bacteria and Archaea, with eukaryotes incorporated among archaea. Eukaryotes represent a small minority of the number of organisms, but, due to their generally much larger size, their collective global biomass is estimated to be about equal to that of prokaryotes. Eukaryotes emerged approximately 2.3–1.8 billion years ago, during the Proterozoic eon, likely as flagellated phagotrophs. Their name comes from the Greek εὖ (''eu'', "well" or "good") and κάρυον (''karyon'', "nut" or "kernel"). Euka ...
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Micelle
A micelle () or micella () (plural micelles or micellae, respectively) is an aggregate (or supramolecular assembly) of surfactant amphipathic lipid molecules dispersed in a liquid, forming a colloidal suspension (also known as associated colloidal system). A typical micelle in water forms an aggregate with the hydrophilic "head" regions in contact with surrounding solvent, sequestering the hydrophobic single-tail regions in the micelle centre. This phase is caused by the packing behavior of single-tail lipids in a bilayer. The difficulty filling all the volume of the interior of a bilayer, while accommodating the area per head group forced on the molecule by the hydration of the lipid head group, leads to the formation of the micelle. This type of micelle is known as a normal-phase micelle (oil-in-water micelle). Inverse micelles have the head groups at the centre with the tails extending out (water-in-oil micelle). Micelles are approximately spherical in shape. Other phases ...
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Complex (chemistry)
A coordination complex consists of a central atom or ion, which is usually metallic and is called the ''coordination centre'', and a surrounding array of bound molecules or ions, that are in turn known as ''ligands'' or complexing agents. Many metal-containing compounds, especially those that include transition metals (elements like titanium that belong to the Periodic Table's d-block), are coordination complexes. Nomenclature and terminology Coordination complexes are so pervasive that their structures and reactions are described in many ways, sometimes confusingly. The atom within a ligand that is bonded to the central metal atom or ion is called the donor atom. In a typical complex, a metal ion is bonded to several donor atoms, which can be the same or different. A polydentate (multiple bonded) ligand is a molecule or ion that bonds to the central atom through several of the ligand's atoms; ligands with 2, 3, 4 or even 6 bonds to the central atom are common. These compl ...
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DNA Condensation
DNA condensation refers to the process of compacting DNA molecules ''in vitro'' or ''in vivo''. Mechanistic details of DNA packing are essential for its functioning in the process of gene regulation in living systems. Condensed DNA often has surprising properties, which one would not predict from classical concepts of dilute solutions. Therefore, DNA condensation ''in vitro'' serves as a model system for many processes of physics, biochemistry and biology. In addition, DNA condensation has many potential applications in medicine and biotechnology. DNA diameter is about 2 nm, while the length of a stretched single molecule may be up to several dozens of centimetres depending on the organism. Many features of the DNA double helix contribute to its large stiffness, including the mechanical properties of the sugar-phosphate backbone, electrostatic repulsion between phosphates (DNA bears on average one elementary negative charge per each 0.17 nm of the double helix), stackin ...
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