Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger
Gene Expression Profiling
Gene Expression
Immunohistochemistry
Gene Expression Regulation
Base Sequence
Molecular Sequence Data
DNA-Directed DNA Polymerase
DNA Primers
Sensitivity and Specificity
RNA Polymerase II
Amino Acid Sequence
DNA
Cloning, Molecular
Transcription, Genetic
Genotype
DNA-Directed RNA Polymerases
Mutation
DNA Polymerase I
Oligonucleotide Probes
Blotting, Southern
Sequence Analysis, DNA
DNA Polymerase II
Multiplex Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
DNA Polymerase III
DNA, Complementary
Electrophoresis, Agar Gel
Gene Amplification
Polymorphism, Genetic
DNA Probes
Alleles
Computer Systems
Escherichia coli
Templates, Genetic
RNA Polymerase I
Cells, Cultured
RNA Polymerase III
Promoter Regions, Genetic
Immunoglobulin Heavy Chains
RNA-Directed DNA Polymerase
DNA Polymerase beta
Exons
Down-regulation of perlecan expression contributes to the inhibition of rat cardiac microvascular endothelial cell proliferation induced by hypoxia. (1/8855)
Exposure of endothelial cells (ECs) to hypoxia leads to a decrease in EC proliferation. However, the mechanism by which hypoxia inhibits EC proliferation is unclear. Perlecan has been reported to play an important role in regulating EC proliferation. We hypothesized that perlecan was involved in the hypoxia-induced inhibition of EC proliferation. To test this hypothesis, rat cardiac microvascular ECs were cultured under normoxic or hypoxic conditions for 12 h and harvested for determination of perlecan mRNA expression using real-time reverse transcription-polymerase chain reaction (RT-PCR). The results showed that exposure of ECs to hypoxia for 12 h induced a decrease in perlecan mRNA expression (61.72%, P<0.05). Concomitantly, the down-regulation of endogenous perlecan induced by hypoxia or the neutralization of endogenous perlecan with anti-perlecan antibody significantly inhibited EC proliferation and responsiveness to basic fibroblast growth factor (bFGF), and decreased focal adhesion kinase (FAK) expression and extracellular signal-regulated kinase 1/2 (ERK1/2) activation. These data indicate that down-regulation of perlecan expression contributes to hypoxia-induced inhibition of rat cardiac microvascular EC proliferation by suppressing FAK-mediated and ERK1/2-dependent growth signals. (+info)Expression of ovarian tumour suppressor OPCML in the female CD-1 mouse reproductive tract. (2/8855)
(+info)Rapid effects of LH on gene expression in the mural granulosa cells of mouse periovulatory follicles. (3/8855)
(+info)Studies of granulosa cell maturation in dominant and subordinate bovine follicles: novel extracellular matrix focimatrix is co-ordinately regulated with cholesterol side-chain cleavage CYP11A1. (4/8855)
(+info)Preterm and infection-driven preterm labor: the role of peroxisome proliferator-activated receptors and retinoid X receptor. (5/8855)
(+info)Gene expression profile of rat ovarian tissue following xenotransplantation into immune-deficient mice. (6/8855)
(+info)Toll-like receptor signaling in hen ovarian granulosa cells is dependent on stage of follicle maturation. (7/8855)
(+info)Temporal regulation of BMP2, BMP6, BMP15, GDF9, BMPR1A, BMPR1B, BMPR2 and TGFBR1 mRNA expression in the oocyte, granulosa and theca cells of developing preovulatory follicles in the pig. (8/8855)
(+info)Polymerase Chain Reaction (PCR) is a laboratory technique used to amplify specific regions of DNA. It enables the production of thousands to millions of copies of a particular DNA sequence in a rapid and efficient manner, making it an essential tool in various fields such as molecular biology, medical diagnostics, forensic science, and research.
The PCR process involves repeated cycles of heating and cooling to separate the DNA strands, allow primers (short sequences of single-stranded DNA) to attach to the target regions, and extend these primers using an enzyme called Taq polymerase, resulting in the exponential amplification of the desired DNA segment.
In a medical context, PCR is often used for detecting and quantifying specific pathogens (viruses, bacteria, fungi, or parasites) in clinical samples, identifying genetic mutations or polymorphisms associated with diseases, monitoring disease progression, and evaluating treatment effectiveness.
Real-Time Polymerase Chain Reaction (RT-PCR) is a laboratory technique used in molecular biology to amplify and detect specific DNA sequences in real-time. It is a sensitive and specific method that allows for the quantification of target nucleic acids, such as DNA or RNA, through the use of fluorescent reporter molecules.
The RT-PCR process involves several steps: first, the template DNA is denatured to separate the double-stranded DNA into single strands. Then, primers (short sequences of DNA) specific to the target sequence are added and allowed to anneal to the template DNA. Next, a heat-stable enzyme called Taq polymerase adds nucleotides to the annealed primers, extending them along the template DNA until a new double-stranded DNA molecule is formed.
During each amplification cycle, fluorescent reporter molecules are added that bind specifically to the newly synthesized DNA. As more and more copies of the target sequence are generated, the amount of fluorescence increases in proportion to the number of copies present. This allows for real-time monitoring of the PCR reaction and quantification of the target nucleic acid.
RT-PCR is commonly used in medical diagnostics, research, and forensics to detect and quantify specific DNA or RNA sequences. It has been widely used in the diagnosis of infectious diseases, genetic disorders, and cancer, as well as in the identification of microbial pathogens and the detection of gene expression.
Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) is a laboratory technique used in molecular biology to amplify and detect specific DNA sequences. This technique is particularly useful for the detection and quantification of RNA viruses, as well as for the analysis of gene expression.
The process involves two main steps: reverse transcription and polymerase chain reaction (PCR). In the first step, reverse transcriptase enzyme is used to convert RNA into complementary DNA (cDNA) by reading the template provided by the RNA molecule. This cDNA then serves as a template for the PCR amplification step.
In the second step, the PCR reaction uses two primers that flank the target DNA sequence and a thermostable polymerase enzyme to repeatedly copy the targeted cDNA sequence. The reaction mixture is heated and cooled in cycles, allowing the primers to anneal to the template, and the polymerase to extend the new strand. This results in exponential amplification of the target DNA sequence, making it possible to detect even small amounts of RNA or cDNA.
RT-PCR is a sensitive and specific technique that has many applications in medical research and diagnostics, including the detection of viruses such as HIV, hepatitis C virus, and SARS-CoV-2 (the virus that causes COVID-19). It can also be used to study gene expression, identify genetic mutations, and diagnose genetic disorders.
Messenger RNA (mRNA) is a type of RNA (ribonucleic acid) that carries genetic information copied from DNA in the form of a series of three-base code "words," each of which specifies a particular amino acid. This information is used by the cell's machinery to construct proteins, a process known as translation. After being transcribed from DNA, mRNA travels out of the nucleus to the ribosomes in the cytoplasm where protein synthesis occurs. Once the protein has been synthesized, the mRNA may be degraded and recycled. Post-transcriptional modifications can also occur to mRNA, such as alternative splicing and addition of a 5' cap and a poly(A) tail, which can affect its stability, localization, and translation efficiency.
Gene expression profiling is a laboratory technique used to measure the activity (expression) of thousands of genes at once. This technique allows researchers and clinicians to identify which genes are turned on or off in a particular cell, tissue, or organism under specific conditions, such as during health, disease, development, or in response to various treatments.
The process typically involves isolating RNA from the cells or tissues of interest, converting it into complementary DNA (cDNA), and then using microarray or high-throughput sequencing technologies to determine which genes are expressed and at what levels. The resulting data can be used to identify patterns of gene expression that are associated with specific biological states or processes, providing valuable insights into the underlying molecular mechanisms of diseases and potential targets for therapeutic intervention.
In recent years, gene expression profiling has become an essential tool in various fields, including cancer research, drug discovery, and personalized medicine, where it is used to identify biomarkers of disease, predict patient outcomes, and guide treatment decisions.
Gene expression is the process by which the information encoded in a gene is used to synthesize a functional gene product, such as a protein or RNA molecule. This process involves several steps: transcription, RNA processing, and translation. During transcription, the genetic information in DNA is copied into a complementary RNA molecule, known as messenger RNA (mRNA). The mRNA then undergoes RNA processing, which includes adding a cap and tail to the mRNA and splicing out non-coding regions called introns. The resulting mature mRNA is then translated into a protein on ribosomes in the cytoplasm through the process of translation.
The regulation of gene expression is a complex and highly controlled process that allows cells to respond to changes in their environment, such as growth factors, hormones, and stress signals. This regulation can occur at various stages of gene expression, including transcriptional activation or repression, RNA processing, mRNA stability, and translation. Dysregulation of gene expression has been implicated in many diseases, including cancer, genetic disorders, and neurological conditions.
Immunohistochemistry (IHC) is a technique used in pathology and laboratory medicine to identify specific proteins or antigens in tissue sections. It combines the principles of immunology and histology to detect the presence and location of these target molecules within cells and tissues. This technique utilizes antibodies that are specific to the protein or antigen of interest, which are then tagged with a detection system such as a chromogen or fluorophore. The stained tissue sections can be examined under a microscope, allowing for the visualization and analysis of the distribution and expression patterns of the target molecule in the context of the tissue architecture. Immunohistochemistry is widely used in diagnostic pathology to help identify various diseases, including cancer, infectious diseases, and immune-mediated disorders.
'Gene expression regulation' refers to the processes that control whether, when, and where a particular gene is expressed, meaning the production of a specific protein or functional RNA encoded by that gene. This complex mechanism can be influenced by various factors such as transcription factors, chromatin remodeling, DNA methylation, non-coding RNAs, and post-transcriptional modifications, among others. Proper regulation of gene expression is crucial for normal cellular function, development, and maintaining homeostasis in living organisms. Dysregulation of gene expression can lead to various diseases, including cancer and genetic disorders.
A base sequence in the context of molecular biology refers to the specific order of nucleotides in a DNA or RNA molecule. In DNA, these nucleotides are adenine (A), guanine (G), cytosine (C), and thymine (T). In RNA, uracil (U) takes the place of thymine. The base sequence contains genetic information that is transcribed into RNA and ultimately translated into proteins. It is the exact order of these bases that determines the genetic code and thus the function of the DNA or RNA molecule.
Molecular sequence data refers to the specific arrangement of molecules, most commonly nucleotides in DNA or RNA, or amino acids in proteins, that make up a biological macromolecule. This data is generated through laboratory techniques such as sequencing, and provides information about the exact order of the constituent molecules. This data is crucial in various fields of biology, including genetics, evolution, and molecular biology, allowing for comparisons between different organisms, identification of genetic variations, and studies of gene function and regulation.
DNA-directed DNA polymerase is a type of enzyme that synthesizes new strands of DNA by adding nucleotides to an existing DNA template in a 5' to 3' direction. These enzymes are essential for DNA replication, repair, and recombination. They require a single-stranded DNA template, a primer with a free 3' hydroxyl group, and the four deoxyribonucleoside triphosphates (dNTPs) as substrates to carry out the polymerization reaction.
DNA polymerases also have proofreading activity, which allows them to correct errors that occur during DNA replication by removing mismatched nucleotides and replacing them with the correct ones. This helps ensure the fidelity of the genetic information passed from one generation to the next.
There are several different types of DNA polymerases, each with specific functions and characteristics. For example, DNA polymerase I is involved in both DNA replication and repair, while DNA polymerase III is the primary enzyme responsible for DNA replication in bacteria. In eukaryotic cells, DNA polymerase alpha, beta, gamma, delta, and epsilon have distinct roles in DNA replication, repair, and maintenance.
DNA primers are short single-stranded DNA molecules that serve as a starting point for DNA synthesis. They are typically used in laboratory techniques such as the polymerase chain reaction (PCR) and DNA sequencing. The primer binds to a complementary sequence on the DNA template through base pairing, providing a free 3'-hydroxyl group for the DNA polymerase enzyme to add nucleotides and synthesize a new strand of DNA. This allows for specific and targeted amplification or analysis of a particular region of interest within a larger DNA molecule.
Sensitivity and specificity are statistical measures used to describe the performance of a diagnostic test or screening tool in identifying true positive and true negative results.
* Sensitivity refers to the proportion of people who have a particular condition (true positives) who are correctly identified by the test. It is also known as the "true positive rate" or "recall." A highly sensitive test will identify most or all of the people with the condition, but may also produce more false positives.
* Specificity refers to the proportion of people who do not have a particular condition (true negatives) who are correctly identified by the test. It is also known as the "true negative rate." A highly specific test will identify most or all of the people without the condition, but may also produce more false negatives.
In medical testing, both sensitivity and specificity are important considerations when evaluating a diagnostic test. High sensitivity is desirable for screening tests that aim to identify as many cases of a condition as possible, while high specificity is desirable for confirmatory tests that aim to rule out the condition in people who do not have it.
It's worth noting that sensitivity and specificity are often influenced by factors such as the prevalence of the condition in the population being tested, the threshold used to define a positive result, and the reliability and validity of the test itself. Therefore, it's important to consider these factors when interpreting the results of a diagnostic test.
RNA Polymerase II is a type of enzyme responsible for transcribing DNA into RNA in eukaryotic cells. It plays a crucial role in the process of gene expression, where the information stored in DNA is used to create proteins. Specifically, RNA Polymerase II transcribes protein-coding genes to produce precursor messenger RNA (pre-mRNA), which is then processed into mature mRNA. This mature mRNA serves as a template for protein synthesis during translation.
RNA Polymerase II has a complex structure, consisting of multiple subunits, and it requires the assistance of various transcription factors and coactivators to initiate and regulate transcription. The enzyme recognizes specific promoter sequences in DNA, unwinds the double-stranded DNA, and synthesizes a complementary RNA strand using one of the unwound DNA strands as a template. This process results in the formation of a nascent RNA molecule that is further processed into mature mRNA for protein synthesis or other functional RNAs involved in gene regulation.
Viral DNA refers to the genetic material present in viruses that consist of DNA as their core component. Deoxyribonucleic acid (DNA) is one of the two types of nucleic acids that are responsible for storing and transmitting genetic information in living organisms. Viruses are infectious agents much smaller than bacteria that can only replicate inside the cells of other organisms, called hosts.
Viral DNA can be double-stranded (dsDNA) or single-stranded (ssDNA), depending on the type of virus. Double-stranded DNA viruses have a genome made up of two complementary strands of DNA, while single-stranded DNA viruses contain only one strand of DNA.
Examples of dsDNA viruses include Adenoviruses, Herpesviruses, and Poxviruses, while ssDNA viruses include Parvoviruses and Circoviruses. Viral DNA plays a crucial role in the replication cycle of the virus, encoding for various proteins necessary for its multiplication and survival within the host cell.
An amino acid sequence is the specific order of amino acids in a protein or peptide molecule, formed by the linking of the amino group (-NH2) of one amino acid to the carboxyl group (-COOH) of another amino acid through a peptide bond. The sequence is determined by the genetic code and is unique to each type of protein or peptide. It plays a crucial role in determining the three-dimensional structure and function of proteins.
Bacterial DNA refers to the genetic material found in bacteria. It is composed of a double-stranded helix containing four nucleotide bases - adenine (A), thymine (T), guanine (G), and cytosine (C) - that are linked together by phosphodiester bonds. The sequence of these bases in the DNA molecule carries the genetic information necessary for the growth, development, and reproduction of bacteria.
Bacterial DNA is circular in most bacterial species, although some have linear chromosomes. In addition to the main chromosome, many bacteria also contain small circular pieces of DNA called plasmids that can carry additional genes and provide resistance to antibiotics or other environmental stressors.
Unlike eukaryotic cells, which have their DNA enclosed within a nucleus, bacterial DNA is present in the cytoplasm of the cell, where it is in direct contact with the cell's metabolic machinery. This allows for rapid gene expression and regulation in response to changing environmental conditions.
Deoxyribonucleic acid (DNA) is the genetic material present in the cells of organisms where it is responsible for the storage and transmission of hereditary information. DNA is a long molecule that consists of two strands coiled together to form a double helix. Each strand is made up of a series of four nucleotide bases - adenine (A), guanine (G), cytosine (C), and thymine (T) - that are linked together by phosphate and sugar groups. The sequence of these bases along the length of the molecule encodes genetic information, with A always pairing with T and C always pairing with G. This base-pairing allows for the replication and transcription of DNA, which are essential processes in the functioning and reproduction of all living organisms.
Molecular cloning is a laboratory technique used to create multiple copies of a specific DNA sequence. This process involves several steps:
1. Isolation: The first step in molecular cloning is to isolate the DNA sequence of interest from the rest of the genomic DNA. This can be done using various methods such as PCR (polymerase chain reaction), restriction enzymes, or hybridization.
2. Vector construction: Once the DNA sequence of interest has been isolated, it must be inserted into a vector, which is a small circular DNA molecule that can replicate independently in a host cell. Common vectors used in molecular cloning include plasmids and phages.
3. Transformation: The constructed vector is then introduced into a host cell, usually a bacterial or yeast cell, through a process called transformation. This can be done using various methods such as electroporation or chemical transformation.
4. Selection: After transformation, the host cells are grown in selective media that allow only those cells containing the vector to grow. This ensures that the DNA sequence of interest has been successfully cloned into the vector.
5. Amplification: Once the host cells have been selected, they can be grown in large quantities to amplify the number of copies of the cloned DNA sequence.
Molecular cloning is a powerful tool in molecular biology and has numerous applications, including the production of recombinant proteins, gene therapy, functional analysis of genes, and genetic engineering.
Genetic transcription is the process by which the information in a strand of DNA is used to create a complementary RNA molecule. This process is the first step in gene expression, where the genetic code in DNA is converted into a form that can be used to produce proteins or functional RNAs.
During transcription, an enzyme called RNA polymerase binds to the DNA template strand and reads the sequence of nucleotide bases. As it moves along the template, it adds complementary RNA nucleotides to the growing RNA chain, creating a single-stranded RNA molecule that is complementary to the DNA template strand. Once transcription is complete, the RNA molecule may undergo further processing before it can be translated into protein or perform its functional role in the cell.
Transcription can be either "constitutive" or "regulated." Constitutive transcription occurs at a relatively constant rate and produces essential proteins that are required for basic cellular functions. Regulated transcription, on the other hand, is subject to control by various intracellular and extracellular signals, allowing cells to respond to changing environmental conditions or developmental cues.
Genotype, in genetics, refers to the complete heritable genetic makeup of an individual organism, including all of its genes. It is the set of instructions contained in an organism's DNA for the development and function of that organism. The genotype is the basis for an individual's inherited traits, and it can be contrasted with an individual's phenotype, which refers to the observable physical or biochemical characteristics of an organism that result from the expression of its genes in combination with environmental influences.
It is important to note that an individual's genotype is not necessarily identical to their genetic sequence. Some genes have multiple forms called alleles, and an individual may inherit different alleles for a given gene from each parent. The combination of alleles that an individual inherits for a particular gene is known as their genotype for that gene.
Understanding an individual's genotype can provide important information about their susceptibility to certain diseases, their response to drugs and other treatments, and their risk of passing on inherited genetic disorders to their offspring.
DNA-directed RNA polymerases are enzymes that synthesize RNA molecules using a DNA template in a process called transcription. These enzymes read the sequence of nucleotides in a DNA molecule and use it as a blueprint to construct a complementary RNA strand.
The RNA polymerase moves along the DNA template, adding ribonucleotides one by one to the growing RNA chain. The synthesis is directional, starting at the promoter region of the DNA and moving towards the terminator region.
In bacteria, there is a single type of RNA polymerase that is responsible for transcribing all types of RNA (mRNA, tRNA, and rRNA). In eukaryotic cells, however, there are three different types of RNA polymerases: RNA polymerase I, II, and III. Each type is responsible for transcribing specific types of RNA.
RNA polymerases play a crucial role in gene expression, as they link the genetic information encoded in DNA to the production of functional proteins. Inhibition or mutation of these enzymes can have significant consequences for cellular function and survival.
A mutation is a permanent change in the DNA sequence of an organism's genome. Mutations can occur spontaneously or be caused by environmental factors such as exposure to radiation, chemicals, or viruses. They may have various effects on the organism, ranging from benign to harmful, depending on where they occur and whether they alter the function of essential proteins. In some cases, mutations can increase an individual's susceptibility to certain diseases or disorders, while in others, they may confer a survival advantage. Mutations are the driving force behind evolution, as they introduce new genetic variability into populations, which can then be acted upon by natural selection.
DNA Polymerase I is a type of enzyme that plays a crucial role in DNA replication and repair in prokaryotic cells, such as bacteria. It is responsible for synthesizing new strands of DNA by adding nucleotides to the 3' end of an existing strand, using the complementary strand as a template.
DNA Polymerase I has several key functions during DNA replication:
1. **5' to 3' exonuclease activity:** It can remove nucleotides from the 5' end of a DNA strand in a process called excision repair, which helps to correct errors that may have occurred during DNA replication.
2. **3' to 5' exonuclease activity:** This enzyme can also proofread newly synthesized DNA by removing incorrect nucleotides from the 3' end of a strand, ensuring accurate replication.
3. **Polymerase activity:** DNA Polymerase I adds new nucleotides to the 3' end of an existing strand, extending the length of the DNA molecule during replication and repair processes.
4. **Pyrophosphorolysis:** It can reverse the polymerization reaction by removing a nucleotide from the 3' end of a DNA strand while releasing pyrophosphate, which is an important step in some DNA repair pathways.
In summary, DNA Polymerase I is a versatile enzyme involved in various aspects of DNA replication and repair, contributing to the maintenance of genetic information in prokaryotic cells.
An oligonucleotide probe is a short, single-stranded DNA or RNA molecule that contains a specific sequence of nucleotides designed to hybridize with a complementary sequence in a target nucleic acid (DNA or RNA). These probes are typically 15-50 nucleotides long and are used in various molecular biology techniques, such as polymerase chain reaction (PCR), DNA sequencing, microarray analysis, and blotting methods.
Oligonucleotide probes can be labeled with various reporter molecules, like fluorescent dyes or radioactive isotopes, to enable the detection of hybridized targets. The high specificity of oligonucleotide probes allows for the precise identification and quantification of target nucleic acids in complex biological samples, making them valuable tools in diagnostic, research, and forensic applications.
Southern blotting is a type of membrane-based blotting technique that is used in molecular biology to detect and locate specific DNA sequences within a DNA sample. This technique is named after its inventor, Edward M. Southern.
In Southern blotting, the DNA sample is first digested with one or more restriction enzymes, which cut the DNA at specific recognition sites. The resulting DNA fragments are then separated based on their size by gel electrophoresis. After separation, the DNA fragments are denatured to convert them into single-stranded DNA and transferred onto a nitrocellulose or nylon membrane.
Once the DNA has been transferred to the membrane, it is hybridized with a labeled probe that is complementary to the sequence of interest. The probe can be labeled with radioactive isotopes, fluorescent dyes, or chemiluminescent compounds. After hybridization, the membrane is washed to remove any unbound probe and then exposed to X-ray film (in the case of radioactive probes) or scanned (in the case of non-radioactive probes) to detect the location of the labeled probe on the membrane.
The position of the labeled probe on the membrane corresponds to the location of the specific DNA sequence within the original DNA sample. Southern blotting is a powerful tool for identifying and characterizing specific DNA sequences, such as those associated with genetic diseases or gene regulation.
DNA Sequence Analysis is the systematic determination of the order of nucleotides in a DNA molecule. It is a critical component of modern molecular biology, genetics, and genetic engineering. The process involves determining the exact order of the four nucleotide bases - adenine (A), guanine (G), cytosine (C), and thymine (T) - in a DNA molecule or fragment. This information is used in various applications such as identifying gene mutations, studying evolutionary relationships, developing molecular markers for breeding, and diagnosing genetic diseases.
The process of DNA Sequence Analysis typically involves several steps, including DNA extraction, PCR amplification (if necessary), purification, sequencing reaction, and electrophoresis. The resulting data is then analyzed using specialized software to determine the exact sequence of nucleotides.
In recent years, high-throughput DNA sequencing technologies have revolutionized the field of genomics, enabling the rapid and cost-effective sequencing of entire genomes. This has led to an explosion of genomic data and new insights into the genetic basis of many diseases and traits.
DNA Polymerase II is a type of enzyme involved in DNA replication and repair in eukaryotic cells. It plays a crucial role in the process of proofreading and correcting errors that may occur during DNA synthesis.
During DNA replication, DNA polymerase II helps to fill in gaps or missing nucleotides behind the main replicative enzyme, DNA Polymerase epsilon. It also plays a significant role in repairing damaged DNA by removing and replacing incorrect or damaged nucleotides.
DNA Polymerase II is highly accurate and has a strong proofreading activity, which allows it to correct most of the errors that occur during DNA synthesis. This enzyme is also involved in the process of translesion synthesis, where it helps to bypass lesions or damage in the DNA template, allowing replication to continue.
Overall, DNA Polymerase II is an essential enzyme for maintaining genomic stability and preventing the accumulation of mutations in eukaryotic cells.
Multiplex polymerase chain reaction (Multiplex PCR) is a laboratory technique that allows the simultaneous amplification and detection of multiple specific DNA sequences in a single reaction. This method utilizes multiple sets of primers, each specifically designed to recognize and bind to a unique target sequence within the DNA sample.
The process involves several steps:
1. Denaturation: The DNA sample is heated to separate the double-stranded DNA into single strands.
2. Annealing: Primers specific to the target sequences are added, and the mixture is cooled, allowing the primers to attach to their respective complementary sequences on the DNA strands.
3. Extension/Amplification: Polymerase enzymes extend the primers along the DNA template, synthesizing new strands of DNA that contain the target sequence. This step is repeated multiple times (usually 25-40 cycles) to exponentially amplify the targeted sequences.
In multiplex PCR, several primer sets are used in a single reaction, allowing for the simultaneous amplification of different target sequences. After amplification, various methods can be employed to distinguish and detect the specific products, such as gel electrophoresis, capillary electrophoresis, or microarray analysis.
Multiplex PCR is widely used in diagnostic tests, pathogen detection, genetic testing, and research applications where multiple DNA targets need to be analyzed simultaneously.
Restriction Fragment Length Polymorphism (RFLP) is a term used in molecular biology and genetics. It refers to the presence of variations in DNA sequences among individuals, which can be detected by restriction enzymes. These enzymes cut DNA at specific sites, creating fragments of different lengths.
In RFLP analysis, DNA is isolated from an individual and treated with a specific restriction enzyme that cuts the DNA at particular recognition sites. The resulting fragments are then separated by size using gel electrophoresis, creating a pattern unique to that individual's DNA. If there are variations in the DNA sequence between individuals, the restriction enzyme may cut the DNA at different sites, leading to differences in the length of the fragments and thus, a different pattern on the gel.
These variations can be used for various purposes, such as identifying individuals, diagnosing genetic diseases, or studying evolutionary relationships between species. However, RFLP analysis has largely been replaced by more modern techniques like polymerase chain reaction (PCR)-based methods and DNA sequencing, which offer higher resolution and throughput.
There doesn't seem to be a specific medical definition for "DNA, protozoan" as it is simply a reference to the DNA found in protozoa. Protozoa are single-celled eukaryotic organisms that can be found in various environments such as soil, water, and the digestive tracts of animals.
Protozoan DNA refers to the genetic material present in these organisms. It is composed of nucleic acids, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), which contain the instructions for the development, growth, and reproduction of the protozoan.
The DNA in protozoa, like in other organisms, is made up of two strands of nucleotides that coil together to form a double helix. The four nucleotide bases that make up protozoan DNA are adenine (A), thymine (T), guanine (G), and cytosine (C). These bases pair with each other to form the rungs of the DNA ladder, with A always pairing with T and G always pairing with C.
The genetic information stored in protozoan DNA is encoded in the sequence of these nucleotide bases. This information is used to synthesize proteins, which are essential for the structure and function of the organism's cells. Protozoan DNA also contains other types of genetic material, such as regulatory sequences that control gene expression and repetitive elements with no known function.
Understanding the DNA of protozoa is important for studying their biology, evolution, and pathogenicity. It can help researchers develop new treatments for protozoan diseases and gain insights into the fundamental principles of genetics and cellular function.
DNA Polymerase III is a critical enzyme in the process of DNA replication in bacteria. It is responsible for synthesizing new strands of DNA by adding nucleotides to the growing chain, based on the template provided by the existing DNA strand. This enzyme has multiple subunits and possesses both polymerase and exonuclease activities. The polymerase activity adds nucleotides to the growing DNA strand, while the exonuclease activity proofreads and corrects any errors that occur during replication. Overall, DNA Polymerase III plays a crucial role in maintaining the accuracy and integrity of genetic information during bacterial cell division.
Complementary DNA (cDNA) is a type of DNA that is synthesized from a single-stranded RNA molecule through the process of reverse transcription. In this process, the enzyme reverse transcriptase uses an RNA molecule as a template to synthesize a complementary DNA strand. The resulting cDNA is therefore complementary to the original RNA molecule and is a copy of its coding sequence, but it does not contain non-coding regions such as introns that are present in genomic DNA.
Complementary DNA is often used in molecular biology research to study gene expression, protein function, and other genetic phenomena. For example, cDNA can be used to create cDNA libraries, which are collections of cloned cDNA fragments that represent the expressed genes in a particular cell type or tissue. These libraries can then be screened for specific genes or gene products of interest. Additionally, cDNA can be used to produce recombinant proteins in heterologous expression systems, allowing researchers to study the structure and function of proteins that may be difficult to express or purify from their native sources.
Electrophoresis, Agar Gel is a laboratory technique used to separate and analyze DNA, RNA, or proteins based on their size and electrical charge. In this method, the sample is mixed with agarose gel, a gelatinous substance derived from seaweed, and then solidified in a horizontal slab-like format. An electric field is applied to the gel, causing the negatively charged DNA or RNA molecules to migrate towards the positive electrode. The smaller molecules move faster through the gel than the larger ones, resulting in their separation based on size. This technique is widely used in molecular biology and genetics research, as well as in diagnostic testing for various genetic disorders.
Gene amplification is a process in molecular biology where a specific gene or set of genes are copied multiple times, leading to an increased number of copies of that gene within the genome. This can occur naturally in cells as a response to various stimuli, such as stress or exposure to certain chemicals, but it can also be induced artificially through laboratory techniques for research purposes.
In cancer biology, gene amplification is often associated with tumor development and progression, where the amplified genes can contribute to increased cell growth, survival, and drug resistance. For example, the overamplification of the HER2/neu gene in breast cancer has been linked to more aggressive tumors and poorer patient outcomes.
In diagnostic and research settings, gene amplification techniques like polymerase chain reaction (PCR) are commonly used to detect and analyze specific genes or genetic sequences of interest. These methods allow researchers to quickly and efficiently generate many copies of a particular DNA sequence, facilitating downstream analysis and detection of low-abundance targets.
Genetic polymorphism refers to the occurrence of multiple forms (called alleles) of a particular gene within a population. These variations in the DNA sequence do not generally affect the function or survival of the organism, but they can contribute to differences in traits among individuals. Genetic polymorphisms can be caused by single nucleotide changes (SNPs), insertions or deletions of DNA segments, or other types of genetic rearrangements. They are important for understanding genetic diversity and evolution, as well as for identifying genetic factors that may contribute to disease susceptibility in humans.
A DNA probe is a single-stranded DNA molecule that contains a specific sequence of nucleotides, and is labeled with a detectable marker such as a radioisotope or a fluorescent dye. It is used in molecular biology to identify and locate a complementary sequence within a sample of DNA. The probe hybridizes (forms a stable double-stranded structure) with its complementary sequence through base pairing, allowing for the detection and analysis of the target DNA. This technique is widely used in various applications such as genetic testing, diagnosis of infectious diseases, and forensic science.
An allele is a variant form of a gene that is located at a specific position on a specific chromosome. Alleles are alternative forms of the same gene that arise by mutation and are found at the same locus or position on homologous chromosomes.
Each person typically inherits two copies of each gene, one from each parent. If the two alleles are identical, a person is said to be homozygous for that trait. If the alleles are different, the person is heterozygous.
For example, the ABO blood group system has three alleles, A, B, and O, which determine a person's blood type. If a person inherits two A alleles, they will have type A blood; if they inherit one A and one B allele, they will have type AB blood; if they inherit two B alleles, they will have type B blood; and if they inherit two O alleles, they will have type O blood.
Alleles can also influence traits such as eye color, hair color, height, and other physical characteristics. Some alleles are dominant, meaning that only one copy of the allele is needed to express the trait, while others are recessive, meaning that two copies of the allele are needed to express the trait.
A viral RNA (ribonucleic acid) is the genetic material found in certain types of viruses, as opposed to viruses that contain DNA (deoxyribonucleic acid). These viruses are known as RNA viruses. The RNA can be single-stranded or double-stranded and can exist as several different forms, such as positive-sense, negative-sense, or ambisense RNA. Upon infecting a host cell, the viral RNA uses the host's cellular machinery to translate the genetic information into proteins, leading to the production of new virus particles and the continuation of the viral life cycle. Examples of human diseases caused by RNA viruses include influenza, COVID-19 (SARS-CoV-2), hepatitis C, and polio.
A computer system is a collection of hardware and software components that work together to perform specific tasks. This includes the physical components such as the central processing unit (CPU), memory, storage devices, and input/output devices, as well as the operating system and application software that run on the hardware. Computer systems can range from small, embedded systems found in appliances and devices, to large, complex networks of interconnected computers used for enterprise-level operations.
In a medical context, computer systems are often used for tasks such as storing and retrieving electronic health records (EHRs), managing patient scheduling and billing, performing diagnostic imaging and analysis, and delivering telemedicine services. These systems must adhere to strict regulatory standards, such as the Health Insurance Portability and Accountability Act (HIPAA) in the United States, to ensure the privacy and security of sensitive medical information.
'Escherichia coli' (E. coli) is a type of gram-negative, facultatively anaerobic, rod-shaped bacterium that commonly inhabits the intestinal tract of humans and warm-blooded animals. It is a member of the family Enterobacteriaceae and one of the most well-studied prokaryotic model organisms in molecular biology.
While most E. coli strains are harmless and even beneficial to their hosts, some serotypes can cause various forms of gastrointestinal and extraintestinal illnesses in humans and animals. These pathogenic strains possess virulence factors that enable them to colonize and damage host tissues, leading to diseases such as diarrhea, urinary tract infections, pneumonia, and sepsis.
E. coli is a versatile organism with remarkable genetic diversity, which allows it to adapt to various environmental niches. It can be found in water, soil, food, and various man-made environments, making it an essential indicator of fecal contamination and a common cause of foodborne illnesses. The study of E. coli has contributed significantly to our understanding of fundamental biological processes, including DNA replication, gene regulation, and protein synthesis.
A genetic template refers to the sequence of DNA or RNA that contains the instructions for the development and function of an organism or any of its components. These templates provide the code for the synthesis of proteins and other functional molecules, and determine many of the inherited traits and characteristics of an individual. In this sense, genetic templates serve as the blueprint for life and are passed down from one generation to the next through the process of reproduction.
In molecular biology, the term "template" is used to describe the strand of DNA or RNA that serves as a guide or pattern for the synthesis of a complementary strand during processes such as transcription and replication. During transcription, the template strand of DNA is transcribed into a complementary RNA molecule, while during replication, each parental DNA strand serves as a template for the synthesis of a new complementary strand.
In genetic engineering and synthetic biology, genetic templates can be manipulated and modified to introduce new functions or alter existing ones in organisms. This is achieved through techniques such as gene editing, where specific sequences in the genetic template are targeted and altered using tools like CRISPR-Cas9. Overall, genetic templates play a crucial role in shaping the structure, function, and evolution of all living organisms.
RNA Polymerase I is a type of enzyme that carries out the transcription of ribosomal RNA (rRNA) genes in eukaryotic cells. These enzymes are responsible for synthesizing the rRNA molecules, which are crucial components of ribosomes, the cellular structures where protein synthesis occurs. RNA Polymerase I is found in the nucleolus, a specialized region within the nucleus of eukaryotic cells, and it primarily transcribes the 5S, 18S, and 28S rRNA genes. The enzyme binds to the promoter regions of these genes and synthesizes the rRNA molecules by adding ribonucleotides in a template-directed manner, using DNA as a template. This process is essential for maintaining normal cellular function and for the production of proteins required for growth, development, and homeostasis.
"Cells, cultured" is a medical term that refers to cells that have been removed from an organism and grown in controlled laboratory conditions outside of the body. This process is called cell culture and it allows scientists to study cells in a more controlled and accessible environment than they would have inside the body. Cultured cells can be derived from a variety of sources, including tissues, organs, or fluids from humans, animals, or cell lines that have been previously established in the laboratory.
Cell culture involves several steps, including isolation of the cells from the tissue, purification and characterization of the cells, and maintenance of the cells in appropriate growth conditions. The cells are typically grown in specialized media that contain nutrients, growth factors, and other components necessary for their survival and proliferation. Cultured cells can be used for a variety of purposes, including basic research, drug development and testing, and production of biological products such as vaccines and gene therapies.
It is important to note that cultured cells may behave differently than they do in the body, and results obtained from cell culture studies may not always translate directly to human physiology or disease. Therefore, it is essential to validate findings from cell culture experiments using additional models and ultimately in clinical trials involving human subjects.
RNA Polymerase III is a type of enzyme that carries out the transcription of DNA into RNA, specifically functioning in the synthesis of small, stable RNAs. These RNAs include 5S rRNA, transfer RNAs (tRNAs), and other small nuclear RNAs (snRNAs). The enzyme recognizes specific promoter sequences in DNA and catalyzes the formation of phosphodiester bonds between ribonucleotides to create a complementary RNA strand. RNA Polymerase III is essential for protein synthesis and cell survival, and its activity is tightly regulated within the cell.
Promoter regions in genetics refer to specific DNA sequences located near the transcription start site of a gene. They serve as binding sites for RNA polymerase and various transcription factors that regulate the initiation of gene transcription. These regulatory elements help control the rate of transcription and, therefore, the level of gene expression. Promoter regions can be composed of different types of sequences, such as the TATA box and CAAT box, and their organization and composition can vary between different genes and species.
In the field of medicine, "time factors" refer to the duration of symptoms or time elapsed since the onset of a medical condition, which can have significant implications for diagnosis and treatment. Understanding time factors is crucial in determining the progression of a disease, evaluating the effectiveness of treatments, and making critical decisions regarding patient care.
For example, in stroke management, "time is brain," meaning that rapid intervention within a specific time frame (usually within 4.5 hours) is essential to administering tissue plasminogen activator (tPA), a clot-busting drug that can minimize brain damage and improve patient outcomes. Similarly, in trauma care, the "golden hour" concept emphasizes the importance of providing definitive care within the first 60 minutes after injury to increase survival rates and reduce morbidity.
Time factors also play a role in monitoring the progression of chronic conditions like diabetes or heart disease, where regular follow-ups and assessments help determine appropriate treatment adjustments and prevent complications. In infectious diseases, time factors are crucial for initiating antibiotic therapy and identifying potential outbreaks to control their spread.
Overall, "time factors" encompass the significance of recognizing and acting promptly in various medical scenarios to optimize patient outcomes and provide effective care.
Immunoglobulin heavy chains are proteins that make up the framework of antibodies, which are Y-shaped immune proteins. These heavy chains, along with light chains, form the antigen-binding sites of an antibody, which recognize and bind to specific foreign substances (antigens) in order to neutralize or remove them from the body.
The heavy chain is composed of a variable region, which contains the antigen-binding site, and constant regions that determine the class and function of the antibody. There are five classes of immunoglobulins (IgA, IgD, IgE, IgG, and IgM) that differ in their heavy chain constant regions and therefore have different functions in the immune response.
Immunoglobulin heavy chains are synthesized by B cells, a type of white blood cell involved in the adaptive immune response. The genetic rearrangement of immunoglobulin heavy chain genes during B cell development results in the production of a vast array of different antibodies with unique antigen-binding sites, allowing for the recognition and elimination of a wide variety of pathogens.
RNA-directed DNA polymerase is a type of enzyme that can synthesize DNA using an RNA molecule as a template. This process is called reverse transcription, and it is the mechanism by which retroviruses, such as HIV, replicate their genetic material. The enzyme responsible for this reaction in retroviruses is called reverse transcriptase.
Reverse transcriptase is an important target for antiretroviral therapy used to treat HIV infection and AIDS. In addition to its role in viral replication, RNA-directed DNA polymerase also has applications in molecular biology research, such as in the production of complementary DNA (cDNA) copies of RNA molecules for use in downstream applications like cloning and sequencing.
A cell line is a culture of cells that are grown in a laboratory for use in research. These cells are usually taken from a single cell or group of cells, and they are able to divide and grow continuously in the lab. Cell lines can come from many different sources, including animals, plants, and humans. They are often used in scientific research to study cellular processes, disease mechanisms, and to test new drugs or treatments. Some common types of human cell lines include HeLa cells (which come from a cancer patient named Henrietta Lacks), HEK293 cells (which come from embryonic kidney cells), and HUVEC cells (which come from umbilical vein endothelial cells). It is important to note that cell lines are not the same as primary cells, which are cells that are taken directly from a living organism and have not been grown in the lab.
DNA polymerase beta is a type of enzyme that plays a crucial role in the repair and maintenance of DNA in cells. It is a member of the DNA polymerase family, which are enzymes responsible for synthesizing new strands of DNA during replication and repair processes.
More specifically, DNA polymerase beta is involved in the base excision repair (BER) pathway, which is a mechanism for correcting damaged or mismatched bases in DNA. This enzyme functions by removing the damaged or incorrect base and replacing it with a new, correct one, using the undamaged strand as a template.
DNA polymerase beta has several key features that make it well-suited to its role in BER. It is highly processive, meaning that it can add many nucleotides to the growing DNA chain before dissociating from the template. It also has a high catalytic rate and is able to efficiently incorporate new nucleotides into the DNA chain.
Overall, DNA polymerase beta is an essential enzyme for maintaining genomic stability and preventing the accumulation of mutations in cells. Defects in this enzyme have been linked to various human diseases, including cancer and neurodegenerative disorders.
Exons are the coding regions of DNA that remain in the mature, processed mRNA after the removal of non-coding intronic sequences during RNA splicing. These exons contain the information necessary to encode proteins, as they specify the sequence of amino acids within a polypeptide chain. The arrangement and order of exons can vary between different genes and even between different versions of the same gene (alternative splicing), allowing for the generation of multiple protein isoforms from a single gene. This complexity in exon structure and usage significantly contributes to the diversity and functionality of the proteome.
Sequence homology in nucleic acids refers to the similarity or identity between the nucleotide sequences of two or more DNA or RNA molecules. It is often used as a measure of biological relationship between genes, organisms, or populations. High sequence homology suggests a recent common ancestry or functional constraint, while low sequence homology may indicate a more distant relationship or different functions.
Nucleic acid sequence homology can be determined by various methods such as pairwise alignment, multiple sequence alignment, and statistical analysis. The degree of homology is typically expressed as a percentage of identical or similar nucleotides in a given window of comparison.
It's important to note that the interpretation of sequence homology depends on the biological context and the evolutionary distance between the sequences compared. Therefore, functional and experimental validation is often necessary to confirm the significance of sequence homology.
Real-time polymerase chain reaction
Reverse transcription polymerase chain reaction
NASBA (molecular biology)
Virus quantification
Lymphogranuloma venereum
W. Ian Lipkin
Primerdesign
Glossary of genetics (M-Z)
Myxomatosis
TaqMan
List of companies of the United Kingdom K-Z
Stephen Bustin
Psittacine beak and feather disease
Microbiome
Isabella Eckerle
MERS
SallyAnn Harbison
Protein detection
Relative fluorescence units
Structural variation
Cancer epigenetics
Shadow effect
Polymerase chain reaction
Luminex Corporation
Beacon designer
Molecular processor
Epizootic hemorrhagic disease virus
Bisulfite sequencing
Ann Marie Kimball
SLC15A2
Real-time polymerase chain reaction - Wikipedia
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Assay for rapid detection1
- Based on individual Flt3-TKD mutations, we designed patient-specific primers to perform a highly sensitive polymerase chain reaction (PCR) assay for rapid detection of minimal residual disease (MRD). (nih.gov)
QPCR6
- A real-time polymerase chain reaction (real-time PCR, or qPCR when used quantitatively) is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR). (wikipedia.org)
- The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines propose that the abbreviation qPCR be used for quantitative real-time PCR and that RT-qPCR be used for reverse transcription-qPCR. (wikipedia.org)
- nonetheless, until now, no validated reference genes from the genus Serratia have been reported that can be used for the normalization of quantitative real-time polymerase chain reaction (RT-qPCR) data. (techscience.com)
- The aim of this study was to utilize the quantitative real-time polymerase chain reaction (qPCR) method for direct quantitative detection of S. aureus in Turkish white cheese samples and identification of the isolated colonies using phenotypic and molecular methods. (iyte.edu.tr)
- One method for doing this is to count the occurrence of certain genetic components, which are proportional to the number of cells present in a cell extract using quantitative, or real time, polymerase chain reaction (qPCR), which is an analytical method used to determine the number of copies of any particular gene per volume tested. (usda.gov)
- Standard-based real-time, or quantitative, polymerase chain reaction (qPCR) quantitation of an unknown sample's DNA concentration (i.e. (usda.gov)
Detection12
- Two common methods for the detection of PCR products in real-time PCR are (1) non-specific fluorescent dyes that intercalate with any double-stranded DNA and (2) sequence-specific DNA probes consisting of oligonucleotides that are labelled with a fluorescent reporter, which permits detection only after hybridization of the probe with its complementary sequence. (wikipedia.org)
- The limit of detection (LODs) of Allplex, PowerChek, and Real-Q was determined by testing the transcribed RNA of SARS-CoV-2 E and the RNA of SARS-CoV Frankfurt1. (frontiersin.org)
- Of the 20 positive samples, the detection rates of positives for Allplex, PowerChek, Real-Q, and StandardM were 90.0, 82.3, 75.0, and 100.0%, respectively, but those of PowerChek and Real-Q would be 100% if out-of-cutoff Cts were counted as positives. (frontiersin.org)
- The existing detection methods for this disease have numerous shortcomings, including low sensitivity, time consuming procedures, and high contamination vulnerability. (medscimonit.com)
- In our study, we developed and evaluated a multiplex real-time polymerase chain reaction (PCR) assay for the detection of C. difficile genes encoding toxin A (tcdA), toxin B (tcdB), and binary toxin (cdtA and cdtB). (psu.edu)
- Real-time PCR relies on the detection and quantitation of amplified DNA in real-time using a fluorescent reporter. (iflybio.com)
- Association Between Rapid Antigen Detection Tests and Real-Time Reverse Transcription-Polymerase Chain Reaction Assay for SARS-CoV-2: A Systematic Review and Meta-Analyses. (bvsalud.org)
- We aimed to assess the association between rapid antigen detection tests and real- time reverse transcription - polymerase chain reaction assay for severe acute respiratory syndrome coronavirus 2 . (bvsalud.org)
- Therefore, primer dimers are an issue in reactions using both probe-based and SYBR Green I dye-based detection. (sigmaaldrich.com)
- PCR: The detection of CMV DNA was performed with polymerase chain reaction (PCR) targeting the viral immediate early 2 (IE-2) region as described by Bopanna et al. (cdc.gov)
- In our previous study, we investigated the prevalence of the BRAF V600 mutation among Indonesian melanoma cases using real-time polymerase chain reaction (RT-PCR) as the detection method and obtained a low percentage compared to Asia and other countries (8). (researchsquare.com)
- Noncompetitive phage anti-immunocomplex real-time polymerase chain reaction for sensitive detection of small molecules. (cdc.gov)
Sensitivity3
- Moreover, a definite diagnosis of epidemic typhus is often delayed because the sensitivity of cell culture and polymerase chain reaction (PCR) methods is low ( 13 ), and serologic diagnosis can be obtained only by using advanced serologic methods such as Western blot analysis after cross-adsorptions. (cdc.gov)
- The use of real-time quantitative RT-PCR to quantify specific mRNAs allows for more rapid testing, higher sensitivity, increased simplicity, and more accuracy. (medscape.com)
- Whenever primer dimer products are produced and amplified, they divert reaction components away from synthesis of the desired product, thereby reducing assay efficiency and sensitivity. (sigmaaldrich.com)
Assays5
- Development and validation of real-time polymerase chain reaction assays specific to four species of Eimeria. (ox.ac.uk)
- The development of quantitative real-time polymerase chain reaction (PCR) assays specific to Eimeria acervulina, Eimeria maxima, Eimeria necatrix and Eimeria tenella is described and validated. (ox.ac.uk)
- The species-specific assays described here, combined with a previously published generic Eimeria species real-time PCR, provide valuable components in a "tool box" to accurately quantify the presence of specific Eimeria species in environmental or within-host phases of the lifecycle with little specialist knowledge. (ox.ac.uk)
- The use of highly reproducible and accurate SYBR-based real-time polymerase chain reaction (PCR) assays instead of performing Taqman-type assays allows low-cost, high-throughput analysis of viral mRNA expression. (biomedcentral.com)
- Dried Blood Spot Real-time Polymerase Chain Reaction Assays to Screen Newborns for Congenital Cytomegalovirus Infection JAMA 2010 202(14) 1375-1382. (cdc.gov)
Methods1
- Materials and Methods Macrophages derived from the THP-1 cell line were differentiated with interleukin-4 to induce a 'wound-healing'-like phenotype, and treated with various dilutions of Arnica m . centesimal (100 times) dilutions (2c, 3c, 5c, 9c, and 15c) or control solvent for 24 hours. (thieme-connect.com)
Immunofluorescence1
- Western blotting, quantitative real-time polymerase chain reaction, immunofluorescence staining, and immunohistochemical staining, were used to evaluate the neuroprotective mechanisms of LIFUS. (aging-us.com)
Multiplex5
- To overcome shortcomings for detecting animal chlamydiosis, a multiplex quantitative polymerase chain reaction (PCR) assay was established for simultaneously detecting and differentiating 3 Chlamydia species ( C. pecorum , C. abortus , and C. psittaci ) by real time PCR based on TaqMan-MGB technology. (medscimonit.com)
- A total of 24 fecal samples (33.80%) were positive for toxigenic C. difficile using either multiplex real-time PCR or culture. (psu.edu)
- Toxin-encoding C. difficile was detected in 23 enriched fecal samples using the multiplex real-time PCR assay and only 15 samples using culture techniques. (psu.edu)
- On the basis of findings of our study, it can be concluded that multiplex real-time PCR carried out on samples enriched for C. difficile is a reliable, sensitive, and specific diagnostic tool for rapid screening and identification of samples contaminated with C. difficile. (psu.edu)
- Similarly, when multiple targets are to be detected simultaneously in multiplex reactions, assay conditions must be optimized to detect all targets equally. (sigmaaldrich.com)
Primers5
- In order to amplify small amounts of DNA, the same methodology is used as in conventional PCR using a DNA template, at least one pair of specific primers, deoxyribonucleotide triphosphates, a suitable buffer solution and a thermo-stable DNA polymerase. (wikipedia.org)
- Initially, DNA is taken from the clinical specimen, as well as certain sequence-specific oligonucleotide primers, thermostable DNA polymerase, nucleotides, and buffer. (medscape.com)
- After the primers bind to the complementary strand of the target DNA, it is extended by DNA polymerase using four deoxynucleotide triphosphate. (iflybio.com)
- To know in detail about designing primers for PCR reactions, click here . (iflybio.com)
- How to design primers for PCR (Polymerase Chain Reaction)? (iflybio.com)
Analyses2
- We present a real-time PCR protocol that can be used for MRD analyses based on individual Flt3-TKD mutations. (nih.gov)
- This study aims to clarify the time-dependent molecular pathways by which TMAO mediates endothelial dysfunction through transcriptomics and metabolomics analyses in human microvascular endothelial cells (HMEC-1). (nature.com)
Genes3
- The standardized real-time PCR assay for toxin genes of C. difficile was used to screen for toxigenic C. difficile in fecal samples from 71 preweaned calves, 53 retail ground meat samples, and 27 pasteurized milk samples. (psu.edu)
- Changes in morphology were imaged, and changes in gene expression of CEC typical genes such as zonula occludens (ZO-1), sodium/potassium (Na/K)-ATPase, paired-like homeodomain 2 (PITX2), and collagen 8 (COL-8) were measured with real-time polymerase chain reaction. (lww.com)
- The expression of selected genes was quantified using real-time polymerase chain reaction. (thejns.org)
Amplify2
- In PCR, a thermostable DNA polymerase is used to amplify target DNA 2-fold with each temperature cycle. (medscape.com)
- These three steps of denaturation, annealing, and extension are repeated in a cyclic manner (20-30 times) in order to amplify the DNA in between the two primer binding sites. (iflybio.com)
Specimens5
- Orthopoxvirus DNA in clinical specimens by real-time PCR. (cdc.gov)
- Here, we report a method of absolute quantitative real-time PCR utilizing SYBR-green fluorescence for the measurement of HPV E7 expression in cervical cytobrush specimens. (biomedcentral.com)
- In order to accomplish this goal, we needed to develop a polymerase chain reaction (PCR) method that would allow us to make direct comparisons between patients in terms of amount of mRNA expressed in cervical specimens. (biomedcentral.com)
- Eighteen cytobrush specimens were analyzed by real-time PCR for the presence of either HPV16 or HPV18 E7 mRNA (Table 1 ). (biomedcentral.com)
- Among participants aged 6-49 years in NHANES between 1999 and 2004 who had stored urine samples available and were cytomegalovirus (CMV) IgG positive, urine specimens were tested with real time PCR to detect CMV. (cdc.gov)
Molecular3
- We also used a model system with MonoMac-6 cells carrying the Flt3-V592A mutation to establish a mutation-specific real-time PCR approach also for this molecular aberration. (nih.gov)
- Interestingly, the molecular signatures were distinct between the two time-points. (nature.com)
- This study demonstrates that TMAO induces distinct time-dependent molecular signatures involving inflammation and remodelling pathways, while pathways such as oxidative stress are also modulated, but in a non-time-dependent manner. (nature.com)
Laboratory3
- This online course is designed for public health and clinical laboratory staff, and persons interested in PCR and real-time PCR techniques. (cdc.gov)
- Polymerase chain reaction (PCR) confirmed laboratory results of suspected cases of cerebrospinal meningitis from January, 2016 to March, 2020 were obtained from the Tamale Public Health Laboratory. (springer.com)
- We analysed national dengue surveillance and laboratory data from 2006 to 2012 by person, place and time. (who.int)
Melting-curve an1
- Subsequently, we developed a method for the measurement of relative allelic expression, by taking advantage of the capability for melting-curve analysis in real-time PCR. (aspetjournals.org)
Method4
- This measurement is made after each amplification cycle, and this is the reason why this method is called real time PCR (that is, immediate or simultaneous PCR). (wikipedia.org)
- We screened for inv22 with our previously reported conventional polymerase chain reaction ( PCR ) method , as well as with a newly developed real-time PCR method . (bvsalud.org)
- With the real-time PCR method , 10 of the severe haemophilia A patients and 3 carriers tested inv22-positive. (bvsalud.org)
- The conventional PCR method and real-time PCR results were comparable in all but one case, where the discrepancy was attributed to sample-specific degradation. (bvsalud.org)
Complementary3
- Reverse transcription is the synthesis of a complementary DNA sequence from an RNA template using reverse transcriptase, which is an RNA-dependent DNA polymerase. (medscape.com)
- [ 1 ] The resultant complementary DNA is amplified by polymerase chain reaction (PCR). (medscape.com)
- Finally, nucleotides complementary to the target DNA are added extending each primer by the thermostable DNA polymerase. (medscape.com)
Amplification1
- It monitors the amplification of a targeted DNA molecule during the PCR (i.e., in real time), not at its end, as in conventional PCR. (wikipedia.org)
Significantly1
- At all timepoints, clinical assessment revealed significantly lower frequencies of positive patch-test reactions to CoNPs compared with CoCl2 or to the positive control. (lu.se)
Periodontitis1
- The purpose of the present investigation is to compare the presence and number of periodontal pathogens in the subgingival microbiota of smokers versus never-smokers with chronic periodontitis and matched probing depths (PDs) using real-time polymerase chain reaction (RT-PCR). (nih.gov)
Quantitative PCR1
- The aim of our study was to develop a real-time quantitative PCR assay by using a species-specific probe that is rapid, sensitive, and specific for detecting R. prowazekii in clinical samples or in body lice in outbreaks of epidemic typhus. (cdc.gov)
Fluorescent1
- The 'threshold line' is the point at which the reaction reaches a fluorescent intensity above background. (ukessays.com)
Experiments1
- RNA samples from cultured cells were analysed by real-time quantitative polymerase chain reaction in five separate experiments. (thieme-connect.com)
Concentration4
- residence time, and increased area under concentration curves (AUC) values. (ajol.info)
- However, the determination of a test sample's DNA concentration assumes that the concentration dependence of the standard and unknown reactions are equivalent. (usda.gov)
- DNA]-unk) assumes that the concentration dependence of the standard and unknown reactions (related to reaction efficiency, E) are equivalent. (usda.gov)
- When significant differences in polymerase E occur, it is not possible to accurately estimate unknown target DNA concentration from the standard solution's slope and intercept (from PCR response, or Ct, versus Log[DNA] data). (usda.gov)
Temperature1
- Hence, Taq polymerase is resistant to high temperatures such as the denaturation temperature. (iflybio.com)
Extract2
- DNA extract solely from whole blood alone is not a suitable specimen for Orthopoxvirus diagnostic testing as the viremic phase may have already passed at the time of rash onset. (cdc.gov)
- This analysis provides: alcohol by volume (ABV), alcohol by weight (ABW), density, apparent extract, real extract, calories, real degree of fermentation (RDF) and apparent degree of fermentation (ADF). (shortsbrewing.com)
Detect1
- Real-time PCR is carried out in a thermal cycler with the capacity to illuminate each sample with a beam of light of at least one specified wavelength and detect the fluorescence emitted by the excited fluorophore. (wikipedia.org)
Rapidly2
- Similar to other bacterial meningitis, the incidence of meningococcal meningitis is seasonally dependent, with peaks during the dry season (December-May) and decreases rapidly, even in times of major epidemics with the start of the rainy season [ 14 , 15 ]. (springer.com)
- At the same time, information about the effects of COVID-19 during pregnancy is evolving rapidly. (degruyter.com)
Endothelial1
- Relative expression of transforming growth factor beta (TGF- β), myosin heavy chain beta (β-MHC), endothelial nitric oxide synthase (eNOS) and glyceraldehydes-3-phosphate dehydrogenase (GAPDH) was studied. (ukessays.com)
Diagnostic1
- Applies to all personnel who perform Orthopoxvirus diagnostic testing by real-time PCR. (cdc.gov)
Introduction1
- Here, the breakthrough for DNA polymerase enzyme that could be used for PCR came with the introduction of Taq polymerase. (iflybio.com)
Principle2
- This basic-level eLearning course provides information on the principle of PCR and real-time PCR. (cdc.gov)
- Its principle is based on the use of DNA polymerase which is an in vitro replication of specific DNA sequences. (intechopen.com)
Fermentation1
- Kefir was sampled during fermentation at seven-time intervals. (biorxiv.org)
Results2
- In resolution WHA60.1, the Health Assembly requested the Director-General to undertake a major review in 2010 of the results of the research undertaken in accordance with the terms of resolution WHA55.15, so that the Sixty-fourth World Health Assembly may reach global consensus on the timing of the destruction of existing variola virus stocks. (who.int)
- Results: Patch testing with CoNPs elicited allergic reactions in Co-sensitized individuals. (lu.se)
Specific2
- We developed a real-time quantitative polymerase chain reaction assay by using a species-specific probe targeting the gltA gene. (cdc.gov)
- Pancreatic cancer is occult without specific symptoms, which means that many patients exhibit locally advanced disease or metastasis at the time of diagnosis [ 1 , 2 ]. (jcancer.org)
Cell1
- The hypoxic regulation of CXCL12 was examined in multiple myeloma plasma cell lines using polymerase chain reaction and western blotting. (haematologica.org)
Development1
- Here, we describe the development of an absolute quantitative real-time PCR assay to quantitate mRNA of the E7 oncogene from HPV 16 and HPV 18 in cervical cytobrush samples. (biomedcentral.com)
Field1
- It is a promising tool for discovery of basic gene expression patterns, determination of drug-induced toxicity in adverse drug reactions, and other uses in the medical field. (researchandmarkets.com)
Standard1
- The gold standard test for identifying SARS-CoV-2, the causative agent of COVID-19, is polymerase chain reaction (PCR). (mdpi.com)
Western1
- The expressions of TLR4, NF-κ B and pro-inflammatory cytokines in the cortical were determined by real time polymerase chain reaction (RT-PCR), western blot, immunohistochemistry, or enzyme-linked immunosorbent assay (ELISA). (karger.com)
Pasteurized milk1
- C. difficile was not detected in ground meat or pasteurized milk by traditional culture or real-time PCR assay. (psu.edu)
Diagnosis2
- The global polymerase chain reaction (PCR) and real-time polymerase chain reaction (PCR) testing market is expected to grow from 7.5 billion in 2019 to about 22.4 billion in 2020 as there is a surge in the number of tests being conducted for the diagnosis of Covid-19. (powershow.com)
- The diagnosis of many infectious diseases, both viral and bacterial, may include the use of reverse transcriptase-polymerase chain reaction (RT-PCR). (medscape.com)
Reverse3
- The acronym "RT-PCR" commonly denotes reverse transcription polymerase chain reaction and not real-time PCR, but not all authors adhere to this convention. (wikipedia.org)
- SARS-CoV-2 real-time reverse-transcription PCR (rRT-PCR) is the most effective testing system currently available to counter COVID-19 epidemics when potent treatments and vaccines are unavailable. (frontiersin.org)
- Since the first SARS-CoV-2 case came to light, real time reverse transcription PCR (rRT-PCR) kits have been approved for use in Korea under Emergency-Use-Authorization (EUA). (frontiersin.org)
Target1
- The target DNA segment is amplified in the range of 10 5 - to 10 6 -fold by repeating this cycle no less than 30-40 times. (medscape.com)
Study2
- Lateral carotid artery aneurysms with immediately induced thrombosis were created in 31 swine for a time-course study. (thejns.org)
- Objectives: The aims of the study were to clarify to what extent CoNPs may elicit ACD responses in participants with Co contact allergy, and to evaluate whether the nanoparticles cause a distinct immune response compared with cobalt chloride (CoCl2) in the skin reactions. (lu.se)