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Plasmid Preparation
A plasmid preparation is a method of DNA extraction and purification for plasmid, plasmid DNA. It is an important step in many molecular biology experiments and is essential for the successful use of plasmids in research and biotechnology. Many methods have been developed to purify plasmid DNA from bacteria. During the purification procedure, the plasmid DNA is often separated from contaminating proteins and genomic DNA. These methods invariably involve three steps: growth of the bacterial culture, harvesting and lysis of the bacteria, and purification of the plasmid DNA. Purification of plasmids is central to molecular cloning. A purified plasmid can be used for many standard applications, such as DNA sequencing, sequencing and transfections into cells. Growth of the bacterial culture Plasmids are almost always purified from liquid Microbiological culture, bacteria cultures, usually ''Escherichia coli, E. coli'', which have been transformation (genetics), transformed and isola ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Plasmid Miniprep
A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from gDNA, chromosomal DNA and can replicate independently. They are most commonly found as small circular, double-stranded DNA molecules in bacteria and archaea; however plasmids are sometimes present in and eukaryote, eukaryotic organisms as well. Plasmids often carry useful genes, such as those involved in antibiotic resistance, virulence, secondary metabolism and bioremediation. While chromosomes are large and contain all the essential genetic information for living under normal conditions, plasmids are usually very small and contain additional genes for special circumstances. Artificial plasmids are widely used as Vector (molecular biology), vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms. In the laboratory, plasmids may be introduced into a cell via transformation (genetics), transformation. Synthetic plasmids are ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Alkaline
In chemistry, an alkali (; from the Arabic word , ) is a basic salt of an alkali metal or an alkaline earth metal. An alkali can also be defined as a base that dissolves in water. A solution of a soluble base has a pH greater than 7.0. The adjective alkaline, and less often, alkalescent, is commonly used in English as a synonym for basic, especially for bases soluble in water. This broad use of the term is likely to have come about because alkalis were the first bases known to obey the Arrhenius definition of a base, and they are still among the most common bases. Etymology The word ''alkali'' is derived from Arabic ''al qalīy'' (or ''alkali''), meaning (see calcination), referring to the original source of alkaline substances. A water-extract of burned plant ashes, called potash and composed mostly of potassium carbonate, was mildly basic. After heating this substance with calcium hydroxide (''slaked lime''), a far more strongly basic substance known as ''caustic ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Peptidoglycan
Peptidoglycan or murein is a unique large macromolecule, a polysaccharide, consisting of sugars and amino acids that forms a mesh-like layer (sacculus) that surrounds the bacterial cytoplasmic membrane. The sugar component consists of alternating residues of β-(1,4) linked N-Acetylglucosamine, ''N''-acetylglucosamine (NAG) and N-Acetylmuramic acid, ''N''-acetylmuramic acid (NAM). Attached to the ''N''-acetylmuramic acid is an oligopeptide chain made of three to five amino acids. The peptide chain can be cross-linked to the peptide chain of another strand forming the 3D mesh-like layer. Peptidoglycan serves a structural role in the bacterial cell wall, giving structural strength, as well as counteracting the osmotic pressure of the cytoplasm. This repetitive linking results in a dense peptidoglycan layer which is critical for maintaining cell form and withstanding high osmotic pressures, and it is regularly replaced by peptidoglycan production. Peptidoglycan hydrolysis and synthesis ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Enzyme
An enzyme () is a protein that acts as a biological catalyst by accelerating chemical reactions. The molecules upon which enzymes may act are called substrate (chemistry), substrates, and the enzyme converts the substrates into different molecules known as product (chemistry), products. Almost all metabolism, metabolic processes in the cell (biology), cell need enzyme catalysis in order to occur at rates fast enough to sustain life. Metabolic pathways depend upon enzymes to catalyze individual steps. The study of enzymes is called ''enzymology'' and the field of pseudoenzyme, pseudoenzyme analysis recognizes that during evolution, some enzymes have lost the ability to carry out biological catalysis, which is often reflected in their amino acid sequences and unusual 'pseudocatalytic' properties. Enzymes are known to catalyze more than 5,000 biochemical reaction types. Other biocatalysts include Ribozyme, catalytic RNA molecules, also called ribozymes. They are sometimes descr ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Lysozyme
Lysozyme (, muramidase, ''N''-acetylmuramide glycanhydrolase; systematic name peptidoglycan ''N''-acetylmuramoylhydrolase) is an antimicrobial enzyme produced by animals that forms part of the innate immune system. It is a glycoside hydrolase that catalyzes the following process: : Hydrolysis of (1→4)-β-linkages between ''N''-acetylmuramic acid and ''N''-acetyl-D-glucosamine residues in a peptidoglycan and between ''N''-acetyl-D-glucosamine residues in chitodextrins Peptidoglycan is the major component of gram-positive bacterial cell wall. This hydrolysis in turn compromises the integrity of bacterial cell walls causing lysis of the bacteria. Lysozyme is abundant in secretions including tears, saliva, human milk, and mucus. It is also present in cytoplasmic granules of the macrophages and the polymorphonuclear neutrophils (PMNs). Large amounts of lysozyme can be found in egg white. C-type lysozymes are closely related to alpha-lactalbumin, α-lactalbumin in sequence and st ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Sonication
image:Sonicator.jpg, A sonicator at the Weizmann Institute of Science during sonicationSonication is the act of applying sound energy to agitate particles in a sample, for various purposes such as the extraction of multiple compounds from plants, microalgae and seaweeds. ultrasound, Ultrasonic frequencies (> 20 kHz) are usually used, leading to the process also being known as ultrasonication or ultra-sonication. In the laboratory, it is usually applied using an ''ultrasonic bath'' or an ''ultrasonic probe'', colloquially known as a ''sonicator''. In a Fourdrinier machine, paper machine, an Ultrasonic foil (papermaking), ultrasonic foil can distribute Cellulose fiber, cellulose fibres more uniformly and strengthen the paper. Effects Sonication has numerous effects, both chemical and physical. The scientific field concerned with understanding the effect of sonic waves on chemical systems is called sonochemistry. The chemical effects of ultrasound do not come from a direc ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Bead Beating
Cell disruption, sometimes referred to as ''digestion'', is a method or process for releasing biological molecules from inside a cell. Methods The production of biologically interesting molecules using cloning and culturing methods allows the study and manufacture of relevant molecules. Except for excreted molecules, cells producing molecules of interest must be disrupted. This page discusses various methods. Another method of disruption is called cell unroofing. Bead method A common laboratory-scale mechanical method for cell disruption uses glass, ceramic, or steel beads, in diameter, mixed with a sample suspended in an aqueous solution. First developed by Tim Hopkins in the late 1970s, the sample and bead mix is subjected to high level agitation by stirring or shaking. Beads collide with the cellular sample, cracking open the cell to release the intracellular components. Unlike some other methods, mechanical shear is moderate during homogenization resulting in excellen ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
French Pressure Cell Press
The French pressure cell press, or French press, is an apparatus used in biological experimentation to disrupt the plasma membrane of cells by passing them through a narrow valve under high pressure. The French press can also be used for disintegration of chloroplasts, homogenates of animal tissue, and other biological particles. It is capable of disrupting cell walls while leaving the cell nucleus undisturbed. The French press was invented by Charles Stacy French of the Carnegie Institution of Washington. The press uses an external hydraulic pump to drive a piston within a larger cylinder that contains the liquid sample. The highly pressurized sample is then squeezed past a needle valve. As the sample passes through the valve, the fluid experiences shear stress and decompression, causing cellular disruption. The major components of a French press are made of stainless steel to prevent sample contamination. A French press is commonly used to break the resilient plasma membra ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Sonication
image:Sonicator.jpg, A sonicator at the Weizmann Institute of Science during sonicationSonication is the act of applying sound energy to agitate particles in a sample, for various purposes such as the extraction of multiple compounds from plants, microalgae and seaweeds. ultrasound, Ultrasonic frequencies (> 20 kHz) are usually used, leading to the process also being known as ultrasonication or ultra-sonication. In the laboratory, it is usually applied using an ''ultrasonic bath'' or an ''ultrasonic probe'', colloquially known as a ''sonicator''. In a Fourdrinier machine, paper machine, an Ultrasonic foil (papermaking), ultrasonic foil can distribute Cellulose fiber, cellulose fibres more uniformly and strengthen the paper. Effects Sonication has numerous effects, both chemical and physical. The scientific field concerned with understanding the effect of sonic waves on chemical systems is called sonochemistry. The chemical effects of ultrasound do not come from a direc ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Chromosomal
A chromosome is a package of DNA containing part or all of the genetic material of an organism. In most chromosomes, the very long thin DNA fibers are coated with nucleosome-forming packaging proteins; in eukaryotic cells, the most important of these proteins are the histones. Aided by chaperone proteins, the histones bind to and condense the DNA molecule to maintain its integrity. These eukaryotic chromosomes display a complex three-dimensional structure that has a significant role in transcriptional regulation. Normally, chromosomes are visible under a light microscope only during the metaphase of cell division, where all chromosomes are aligned in the center of the cell in their condensed form. Before this stage occurs, each chromosome is duplicated (S phase), and the two copies are joined by a centromere—resulting in either an X-shaped structure if the centromere is located equatorially, or a two-armed structure if the centromere is located distally; the joined co ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Supercoiled
DNA supercoiling refers to the amount of twist in a particular DNA strand, which determines the amount of strain on it. A given strand may be "positively supercoiled" or "negatively supercoiled" (more or less tightly wound). The amount of a strand's supercoiling affects a number of biological processes, such as compacting DNA and regulating access to the genetic code (which strongly affects Nucleic acid metabolism, DNA metabolism and possibly gene expression). Certain enzymes, such as topoisomerases, change the amount of DNA supercoiling to facilitate functions such as DNA replication and Transcription (genetics), transcription. The amount of supercoiling in a given strand is described by a mathematical formula that compares it to a reference state known as "relaxed B-form" DNA. Overview In a "relaxed" Nucleic acid double helix, double-helical segment of B-DNA, the two strands twist around the helical axis once every 10.4–10.5 base pairs of DNA sequence, sequence. Adding or ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Buffer Solution
A buffer solution is a solution where the pH does not change significantly on dilution or if an acid or base is added at constant temperature. Its pH changes very little when a small amount of strong acid or base is added to it. Buffer solutions are used as a means of keeping pH at a nearly constant value in a wide variety of chemical applications. In nature, there are many living systems that use buffering for pH regulation. For example, the bicarbonate buffering system is used to regulate the pH of blood, and bicarbonate also acts as a buffer in the ocean. Principles of buffering Buffer solutions resist pH change because of a chemical equilibrium between the weak acid HA and its conjugate base A−: When some strong acid is added to an equilibrium mixture of the weak acid and its conjugate base, hydrogen ions (H+) are added, and the equilibrium is shifted to the left, in accordance with Le Chatelier's principle. Because of this, the hydrogen ion concentration increas ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
