Polymerase chain reaction. Medical search. FAQ

subunit
This gene encodes a DNA-directed RNA polymerase I subunit. (nih.gov)
IPB001572 (RNA polymerases beta subunit) with a combined E-value of 3.4e-230. (northwestern.edu)

sequences
Improvements to the in situ polymerase chain reaction (PCR), a process of in vitro enzymatic amplification of specific nucleic acid sequences within the cells where they originate, can be achieved by changing the way that the enzymatic reaction is started. (google.es)
Multiplex polymerase chain reaction (Multiplex PCR) refers to the use of polymerase chain reaction to amplify several different DNA sequences simultaneously (as if performing many separate PCR reactions all together in one reaction). (wikipedia.org)

transcription
DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. (nih.gov)

process
This process amplifies DNA in samples using multiple primers and a temperature-mediated DNA polymerase in a thermal cycler. (wikipedia.org)

start
Reaction initiation is delayed until the start. (google.es)

Amplification
Thermocyclers provide tight control over both the reaction temperature and the duration of each temperature step, ensuring efficient amplification. (neb.com)
In particular, the compact unit can be used to: (1) take cell samples, (2) extract the DNA, (3) perform polymerase chain reaction (PCR) amplification to generate copies of the DNA, (4) perform electrophoresis to measure the spacing between DNA bands (to create the genetic fingerprint), and (5) perform short tandem repeat (STR) analysis to create a unique genetic profile for the individual. (medgadget.com)
Improvements to the in situ polymerase chain reaction (PCR), a process of in vitro enzymatic amplification of specific nucleic acid sequences within the cells where they originate, can be achieved by changing the way that the enzymatic reaction is started. (google.es)
The in vitro amplification of genomic or complementary deoxyribonucleic acid (cDNA) target sequences by polymerase chain reaction has opened new avenues toward the detection of minimal residual leukemic cells at frequencies of 1:10000 to 1:1000000. (springer.com)
Hansen-Hagge TE, Yokota S, Bartram CR (1989) Detection of minimal residual disease in acute lymphoblastic leukemia by in vitro amplification of rearranged T-cell receptor δ chain sequences. (springer.com)
PCR inhibitors are any factor which prevent the amplification of nucleic acids through the polymerase chain reaction (PCR). (wikipedia.org)
By comparing the amplification of this template in the mixture to the amplification observed in a separate experiment in which the same template is used in the absence of inhibitors, the extent of inhibition in the investigated reaction mixture can be inferred. (wikipedia.org)
Nonspecific primer binding obscures polymerase chain reaction results, as the nonspecific sequences to which primers anneal in early steps of amplification will "swamp out" any specific sequences because of the exponential nature of polymerase amplification. (wikipedia.org)
Nested polymerase chain reaction (Nested PCR) is a modification of polymerase chain reaction intended to reduce non-specific binding in products due to the amplification of unexpected primer binding sites. (wikipedia.org)
The PCR method is extremely sensitive, requiring only a few DNA molecules in a single reaction for amplification across several orders of magnitude. (wikipedia.org)
Several "high-fidelity" DNA polymerases, having engineered 3' to 5' exonuclease activity, have become available that permit more accurate amplification for use in PCRs for sequencing or cloning of products. (wikipedia.org)
Although similar to PCR in using repeated applications of DNA polymerase, the process they usually describe employs just a single primer-template complex, and therefore would not lead to the exponential amplification seen in PCR. (wikipedia.org)
In the one-step approach, the entire reaction from cDNA synthesis to PCR amplification occurs in a single tube. (wikipedia.org)
On the other hand, the two-step reaction requires that the reverse transcriptase reaction and PCR amplification be performed in separate tubes. (wikipedia.org)
Template-switching polymerase chain reaction (TS-PCR) is a method of reverse transcription and polymerase chain reaction (PCR) amplification that relies on a natural PCR primer sequence at the polyadenylation site and adds a second primer through the activity of murine leukemia virus reverse transcriptase. (wikipedia.org)

Primers
During a typical PCR, template DNA (containing the region of interest) is mixed with deoxynucleotides (dNTPs), a DNA polymerase and primers. (neb.com)
Primers are short segments of DNA that are complementary with the template DNA upstream of the region of interest and serve as recruitment sites for the polymerase. (neb.com)
Primers bind to the strand to give the polymerase something to extend. (physicsforums.com)
Rychlik W (1993) Selection of primers for polymerase chain reaction. (els.net)
The touchdown polymerase chain reaction or touchdown style polymerase chain reaction is a method of polymerase chain reaction by which primers avoid amplifying nonspecific sequences. (wikipedia.org)
Nested polymerase chain reaction involves two sets of primers, used in two successive runs of polymerase chain reaction, the second set intended to amplify a secondary target within the first run product. (wikipedia.org)
The target DNA undergoes the first run of polymerase chain reaction with the first set of primers, shown in green. (wikipedia.org)
The product from the first reaction undergoes a second run with the second set of primers, shown in red. (wikipedia.org)
A PCR solution is made similarly to a TaqMan assay, which consists of template DNA (or RNA), fluorescence-quencher probes, primers, and a PCR master mix, which contains DNA polymerase, dNTPs, MgCl2, and reaction buffers at optimal concentrations. (wikipedia.org)
This process amplifies DNA in samples using multiple primers and a temperature-mediated DNA polymerase in a thermal cycler. (wikipedia.org)
Secondary structures in the DNA can result in folding or knotting of DNA template or primers, leading to decreased product yield or failure of the reaction. (wikipedia.org)
Primers which bind to incorrect template sites are stabilized in the presence of excessive magnesium concentrations and so results in decreased specificity of the reaction. (wikipedia.org)
Sequence-specific oligonucleotides were used both as building blocks for the gene, and as primers and templates for DNA polymerase. (wikipedia.org)
By 1971 researchers in Khorana's project, concerned over their yields of DNA, began looking at "repair synthesis" - an artificial system of primers and templates that allows DNA polymerase to copy segments of the gene they are synthesizing. (wikipedia.org)
The use of DNA polymerase to extend oligonucleotide primers was a common procedure in DNA sequencing and the production of cDNA for cloning and expression. (wikipedia.org)
As in most PCR reactions, two primers-one for each end-are used per sequence. (wikipedia.org)
Following separation, the eluted fragments of appropriate size are subject to normal PCR, using the outermost primers used in the initial, mutagenic PCR reactions. (wikipedia.org)
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, deoxyribonucleotides, a suitable buffer solution and a thermo-stable DNA polymerase. (wikipedia.org)
Polymerase chain reaction is done by placing a mixture of the desired DNA, DNA polymerase, primers, and nucleotide bases into a machine. (wikipedia.org)

thermostable DNA polymerase
citation needed] Magnesium is required as a co-factor for thermostable DNA polymerase. (wikipedia.org)

Application of Polymerase Chain Re
In the following we will briefly summarize our recent experience with the application of polymerase chain reaction (PCR) strategies to patients with chronic myelocytic leukemia (CML) and acute lymphoblastic leukemia (ALL) in complete remission according to clinical and laboratory parameters. (springer.com)

amplify
The polymerase chain reaction (PCR) is a method to rapidly amplify sequences of DNA. (neb.com)
PCR is a series of temperature-controlled reactions which enable us to amplify a very tiny sample of DNA, producing enough material for it to be analysed or used in DNA profiling. (abpischools.org.uk)
Inverse polymerase chain reaction (Inverse PCR) is a variant of the polymerase chain reaction that is used to amplify DNA with only one known sequence. (wikipedia.org)
If the primer initially (during the higher-temperature phases) binds to the sequence of interest, subsequent rounds of polymerase chain reaction can be performed upon the product to further amplify those fragments. (wikipedia.org)
Polymerase chain reaction itself is the process used to amplify DNA samples, via a temperature-mediated DNA polymerase. (wikipedia.org)
Digital polymerase chain reaction (digital PCR, DigitalPCR, dPCR, or dePCR) is a biotechnological refinement of conventional polymerase chain reaction methods that can be used to directly quantify and clonally amplify nucleic acids strands including DNA, cDNA or RNA. (wikipedia.org)
Multiplex polymerase chain reaction (Multiplex PCR) refers to the use of polymerase chain reaction to amplify several different DNA sequences simultaneously (as if performing many separate PCR reactions all together in one reaction). (wikipedia.org)
It is possible to amplify larger pieces of up to 50,000 base pairs with a slower heating cycle and special polymerases. (wikipedia.org)
In 1986, Saiki started to use Thermophilus aquaticus (Taq) DNA polymerase to amplify segments of DNA. (wikipedia.org)

qPCR
RT-PCR is often confused with real-time polymerase chain reaction (qPCR) by students and scientists alike, but they are separate and distinct techniques. (wikipedia.org)
A real-time polymerase chain reaction (Real-Time PCR), also known as quantitative polymerase chain reaction (qPCR), is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR). (wikipedia.org)

sequence
DNA polymerases, whether from humans, bacteria, or viruses, cannot copy a chain of DNA without a short sequence of nucleotides to "prime" the process, or get it started. (accessexcellence.org)
A PCR vial contains all the necessary components for DNA duplication: a piece of DNA, large quantities of the four nucleotides, large quantities of the primer sequence, and DNA polymerase. (accessexcellence.org)
Polymerase chain reaction (PCR) is a revolutionary laboratory technique that enables the replication of a specific DNA sequence. (news-medical.net)
This polymerase chain reaction/sequence specific primer (PCR-SSP) method gave perfect correlation with serological typing on 40 individuals of previously determined HLA type and on 50 further clinical samples elaluated blind. (hindawi.com)
Of course, if any part of the inhibition occurring in the sample-derived reaction mixture is sequence-specific, then this method will yield an underestimate of the inhibition as it applies to the investigate sequence(s). (wikipedia.org)
Then, like other polymerase chain reaction processes, the DNA is amplified by the temperature-sensitive DNA polymerase: A target region with an internal section of known sequence and unknown flanking regions is identified Genomic DNA is digested into fragments of a few kilobases by a usually low-moderate frequency (6-8 base) cutting restriction enzyme. (wikipedia.org)
The technique employed an oligonucleotide primer, DNA polymerase, and modified nucleotide precursors that block further extension of the primer in sequence-dependent manner. (wikipedia.org)

enzyme
Taq DNA Polymerase: This is a thermal stable enzyme isolated from thermophilic bacteria. (slideshare.net)
The polymerase chain reaction method is used to quantify nucleic acids by amplifying a nucleic acid molecule with the enzyme DNA polymerase. (wikipedia.org)
Taq polymerase is a magnesium-dependent enzyme and determining the optimum concentration to use is critical to the success of the PCR reaction. (wikipedia.org)
Some of the components of the reaction mixture such as template concentration, dNTPs and the presence of chelating agents (EDTA) or proteins can reduce the amount of free magnesium present thus reducing the activity of the enzyme. (wikipedia.org)
active site The part of an enzyme or antibody where the chemical reaction occurs. (wikipedia.org)
His methods for modeling enzyme catalyzed reactions and reactions inside cells are one of the fast moving frontiers in computational biology according to the Institute for Scientific Information. (wikipedia.org)
He has also focused his research attention on deriving mathematical expressions to estimate enzyme kinetics parameters under different reaction conditions. (wikipedia.org)
He has systematically obtained equations to estimate kinetic parameters for the family of Michaelis-Menten reaction mechanisms and determined their region of validity for the initial enzyme and substrate concentrations. (wikipedia.org)
Dr. Schnell derived a generic expression, known nowadays as the Schnell-Mendoza equation, to determine the Michaelis-Menten constant and maximum velocity for enzyme catalyzed reactions following Michaelis-Menten kinetics using time course data. (wikipedia.org)

nucleic
9. A method in accordance with claim 1 wherein said first subset of PCR reagents consists of all PCR reagents except a nucleic acid polymerase. (google.es)

nucleotide bases
To copy DNA, polymerase requires two other components: a supply of the four nucleotide bases and something called a primer. (accessexcellence.org)

Mullis
Recently, the application of the polymerase chain reaction (peR) technique developed by K. D. Mullis and detailed in the study by Saiki et al. (springer.com)
These early precursors to PCR were carefully scrutinized in a patent lawsuit, and are discussed in Mullis' chapters in The Polymerase Chain Reaction (1994). (wikipedia.org)
it was there, in 1983, that Mullis invented his prize-winning improvements to the polymerase chain reaction. (wikipedia.org)

Nucleotides
The Taq polymerase begins adding nucleotides to the primer and eventually makes a complementary copy of the template. (accessexcellence.org)

specificity of the reaction
Touchdown increases specificity of the reaction at higher temperatures and increases the efficiency towards the end by lowering the annealing temperature. (wikipedia.org)

Replication
DNA polymerase will be added to complete the process of repair replication. (wikipedia.org)
The replication reaction continues to produce a fully dimerised DNA fragment. (wikipedia.org)
A complication at that point was that the DNA polymerase used was destroyed by the high heat used at the start of each replication cycle and had to be replaced. (wikipedia.org)

template
As the two strands separate, DNA polymerase makes a copy using each strand as a template. (accessexcellence.org)
If the template contains a G, it adds a C to the new chain, and so on to the end of the DNA strand. (accessexcellence.org)
The three steps in the polymerase chain reaction - the separation of the strands, annealing the primer to the template, and the synthesis of new strands - take less than two minutes. (accessexcellence.org)
Vogelstein diluted the patient samples such that they expected an average of 1 template molecule per 2 wells of a reaction plate. (sigmaaldrich.com)
Detection of bifidobacteria can be achieved using DNA extracted from human faeces as template in PCR reactions. (scribd.com)
In order to try to assess the extent of inhibition that occurs in a reaction, a control can be performed by adding a known amount of a template to the investigated reaction mixture (based on the sample under analysis). (wikipedia.org)
PCR is routinely assessed against a negative control reaction that is set up identically to the experimental PCR, but without template DNA, and performed alongside the experimental PCR. (wikipedia.org)

reagents
These chips have integrated chambers and valves for partitioning samples and reaction reagents. (sigmaaldrich.com)
One lab area is dedicated to preparation and handling of pre-PCR reagents and the setup of the PCR reaction, and another area to post-PCR processing, such as gel electrophoresis or PCR product purification. (wikipedia.org)

Optimization
White, Bruce A(Sep 2005) Polymerase Chain Reaction (PCR): Design and Optimization of Reactions. (els.net)
The polymerase chain reaction (PCR) is a commonly used molecular biology tool for amplifying DNA, and various techniques for PCR optimization which have been developed by molecular biologists to improve PCR performance and minimize failure. (wikipedia.org)

strands
3) Finally, the DNA-Polymerase has to fill in the missing strands. (slideshare.net)

molecular biology
Reverse transcription polymerase chain reaction (RT-PCR), a variant of polymerase chain reaction (PCR), is a technique commonly used in molecular biology to detect RNA expression. (wikipedia.org)
Molecular biology consists of different techniques including Polymerase chain reaction, Gel electrophoresis, and macromolecule blotting to manipulate DNA. (wikipedia.org)

genetic
To order a free set of ABPI schools posters on the following topics: biotechnology, cloning, genetic engineering, unravelling the genome, polymerase chain reaction and stem cells, please fill in our order form . (abpischools.org.uk)

enzymatic
The one-step approach is thought to minimize experimental variation by containing all of the enzymatic reactions in a single environment. (wikipedia.org)

quantitative
His work has focused to resolve the ambiguities in the quantitative analysis and modeling of reactions inside cells. (wikipedia.org)

samples
As well as methods for the removal of inhibitors from samples before PCR, some DNA polymerases offer varying resistance to different inhibitors and increasing the concentration of the chosen DNA polymerase also confers some resistance to polymerase-targeted inhibitors. (wikipedia.org)

copies
Taking into account the time it takes to change the temperature of the reaction vial, 1 million copies can be ready in about three hours. (accessexcellence.org)
When using dPCR, the sample is diluted and separated into a large number of reaction chambers, such that each partition contains either one or no copies of target. (sigmaaldrich.com)

Reverse
a laboratory technology known as reverse transcription-polymerase chain reaction (RT-PCR), a powerful tool used in research and in the diagnosis of diseases such as cancer. (britannica.com)
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)

Diagnosis
This study aimed to establish an accurate and sensitive polymerase chain reaction [‎PCR]‎ technique for the diagnosis of active human brucellosis in Egypt. (who.int)

strand
During the extension step (typically 68-72°C) the polymerase extends the primer to form a nascent DNA strand. (neb.com)
In a PCR reaction DNA polymerase is responsible for building the new strand of DNA. (physicsforums.com)
However because of the mechanism by which DNA polymerase works it cannot just build a new strand opposite the old, it can only build off of existing DNA. (physicsforums.com)

step of the reaction
This is a machine that heats and cools the reaction tubes within it to the precise temperature required for each step of the reaction. (slideshare.net)
The final step of the reaction is to make a complete copy of the templates. (accessexcellence.org)

cycles
The amount of product from the PCR increases with the number of temperature cycles that the reaction is subjected to. (wikipedia.org)

variant
The overlap extension polymerase chain reaction (or OE-PCR) is a variant of PCR. (wikipedia.org)

resistance
Other valuable properties of the chimeric polymerases TopoTaq and PfuC2 include enhanced thermostability, specificity and resistance to contaminants and inhibitors. (wikipedia.org)

concentration
When using this technique, the sample is taken to limiting dilution and the number of positive and negative reactions is used to determine a precise measurement of target concentration. (sigmaaldrich.com)
Essential aspects of binding assays include, but are not limited to, the concentration level of reactants or products (see radioactive section), maintaining the equilibrium constant of reactants throughout the assay, and the reliability and validity of linked reactions. (wikipedia.org)

stochastic
However, above this size, product yields often decrease, as with increasing length stochastic effects such as premature termination by the polymerase begin to affect the efficiency of the PCR. (wikipedia.org)
Stochastic approaches for modelling in vivo reactions. (wikipedia.org)

technique
DNA-based procedures are becoming increasingly common within the analytical laboratory where the polymerase chain reaction (PCR) has become an indispensable technique. (rsc.org)
In recognition of his invention of the polymerase chain reaction (PCR) technique, he shared the 1993 Nobel Prize in Chemistry with Michael Smith and earned the Japan Prize in the same year. (wikipedia.org)
The Taq polymerase was heat resistant and would only need to be added once, thus making the technique dramatically more affordable and subject to automation. (wikipedia.org)

dPCR
dPCR also carries out a single reaction within a sample, however the sample is separated into a large number of partitions and the reaction is carried out in each partition individually. (wikipedia.org)
dPCR improves upon the current PCR practices by dividing up the reaction into multiple, smaller reactions. (wikipedia.org)

binding protein
These are polymerases fused to a processivity-enhancing DNA-binding protein, enhancing adherence of the polymerase to the DNA. (wikipedia.org)

Applications
Lukyanov S.A., Lukyanov K.A., Gurskaya N.G., Bogdanova E.A., Buzdin A.A. (2007) Selective Suppression of Polymerase Chain Reaction and Its Most Popular Applications. (springer.com)

partitions
The number of reaction chambers or partitions varies between systems, from several thousand when using the QX100 Bio-Rad system to millions when using the RainDrop approach. (sigmaaldrich.com)

works
Since the Taq polymerase works best at around 75 degrees C (the temperature of the hot springs where the bacterium was discovered), the temperature of the vial is raised. (accessexcellence.org)
This overview will walk you through how the Polymerase Chain Reaction (PCR) works. (neb.com)

single
PCR carries out one reaction per single sample. (wikipedia.org)
Annealing temperatures for each of the primer sets must be optimized to work correctly within a single reaction, and amplicon sizes, i.e., their base pair length, should be different enough to form distinct bands when visualized by gel electrophoresis. (wikipedia.org)

different
The three parts of the polymerase chain reaction are carried out in the same vial, but at different temperatures. (accessexcellence.org)

extension
Only the duplex without overlap at the 5' end will allow extension by DNA polymerase in 3' to 5' direction. (wikipedia.org)

temperatures
The polymerase chain reaction, now widely used in research laboratories and doctor's offices, relies on the ability of DNA-copying enzymes to remain stable at high temperatures. (accessexcellence.org)
The earliest steps of a touchdown polymerase chain reaction cycle have high annealing temperatures. (wikipedia.org)