The diagnosis of many infectious diseases, both viral and bacterial, may include the use of reverse transcriptase-polymerase chain reaction (RT-PCR). (medscape.com)
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)
The same sequence of reactions is widely used in the laboratory to convert RNA to DNA for use in molecular cloning, RNA sequencing, polymerase chain reaction (PCR), or genome analysis. (wikipedia.org)
In virus species with reverse transcriptase lacking DNA-dependent DNA polymerase activity, creation of double-stranded DNA can possibly be done by host-encoded DNA polymerase δ, mistaking the viral DNA-RNA for a primer and synthesizing a double-stranded DNA by a similar mechanism as in primer removal, where the newly synthesized DNA displaces the original RNA template. (wikipedia.org)
This research was aimed to detect Newcastle Disease virus (NDV) in chickens suspected with ND using the Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) technique. (ugm.ac.id)
Polymerase chain reaction (PCR) was invented by Mullis in 1983 and patented in 1985. (intechopen.com)
Is the Subject Area "Reverse transcriptase-polymerase chain reaction" applicable to this article? (plos.org)
In a single tube, RNA is first converted to cDNA by a reverse transcriptase, and then a DNA-dependent DNA polymerase amplifies the cDNA, enabling quantitation via qPCR. (neb.com)
In the Luna Probe One-Step RT-qPCR Kit (No ROX), Hot Start Taq DNA Polymerase is combined with a novel WarmStart-activated reverse transcriptase, allowing dual control of enzyme activity via reversible, aptamer-based inhibition. (neb.com)
Next-generation sequencing analyses revealed the expression of TRPV1 in several native breast cancer tissues, which was subsequently validated via reverse transcriptase-polymerase chain reaction. (dovepress.com)
Using semi-quantitative reverse-transcription polymerase chain reaction (RT-PCR) and western blots experiments, we compared changes in ETS-1 and ETS-2 expression, their protein levels, and the regulation of some of their target gene expressions at different stages of the ocular tumoral progression in the transgenic mouse model, Tyrp-1-TAg, with those in normal eyes from control mice of the same age. (molvis.org)
The methylation specific polymerase chain reaction in conjunction with sequencing verification was used to establish the methylation patterns of the 14 genes in the liver tissues of four healthy liver donors, as well as tumor and the paired non-cancerous tissues of 30 HCC patients. (wjgnet.com)
Vomitus and stool samples from affected passengers were tested for norovirus by reverse transcriptase-polymerase chain reaction. (cdc.gov)
Hepatitis C RNA: Testing for HCV RNA by reverse-transcriptase ¬polymerase-chain reaction (RT-PCR) amplification of the 5' noncoding region was performed on anti-HCV positive samples. (cdc.gov)
RocketScriptTM Reverse Transcriptase is an independently developed thermostable reverse transcriptase derived from M-MLV. (thomassci.com)
Simplified procedure by including all components necessary for cDNA synthesis such as Thermostable Reverse Transcriptase, RNase inhibitor, etc. (thomassci.com)
Murine leuke3
Reverse transcriptases were discovered by Howard Temin at the University of Wisconsin-Madison in Rous sarcoma virions and independently isolated by David Baltimore in 1970 at MIT from two RNA tumour viruses: murine leukemia virus and again Rous sarcoma virus. (wikipedia.org)
This is achieved through the use of a reverse transcriptase such as AMV RT (avian myeloblastis virus reverse transcriptase) or M-MuLV RT (moloney murine leukemia virus reverse transcriptase). (sigmaaldrich.com)
Enzyme8
A reverse transcriptase (RT) is an enzyme used to generate complementary DNA (cDNA) from an RNA template, a process termed reverse transcription. (wikipedia.org)
Reverse-transcribing RNA viruses, such as retroviruses, use the enzyme to reverse-transcribe their RNA genomes into DNA, which is then integrated into the host genome and replicated along with it. (wikipedia.org)
A domain on the reverse transcriptase enzyme called RNAse H degrades the U5 and R regions on the 5' end of the RNA. (wikipedia.org)
Five heterozygous, nonsynonymous mutations (which cause an amino acid change in the corresponding protein) were identified in TERT, the gene for the telomerase reverse transcriptase catalytic enzyme, among seven unrelated patients. (nih.gov)
The higher thermostability of this enzyme compared to M-MuLV RT allows the reaction to be performed at +42°C, giving higher specificity and better resolution of secondary structures compared to a reaction performed at +37°C. (sigmaaldrich.com)
Nucleoside reverse transcriptase inhibitors (NRTIs) such as AZT (zidovudine)-the first drug approved by the U.S. Food and Drug Administration to prolong the lives of AIDS patients-act by terminating the proviral DNA chain before the enzyme can finish transcription. (britannica.com)
NRTIs are often given in combination with non-nucleoside reverse transcriptase inhibitors (NNRTIs) such as efavirenz that act by binding to and altering the shape of the enzyme itself, thereby blocking the enzyme's function. (britannica.com)
The Luna WarmStart RT Enzyme Mix is supplied at 20X concentration and contains Luna WarmStart Reverse Transcriptase as well as Murine RNase Inhibitor to aid in preventing RNA degradation (see also template preparation in product manual). (neb.com)
Synthesis7
The First Strand cDNA Synthesis Kit is used for the synthesis of the first strand cDNA as the starting reaction for two-step RT PCR.The kit includes Reverse Transcriptase AMV for first strand synthesis, two different primers, our PCR Nucleotide Mix, and Control Neo pa RNA. (sigmaaldrich.com)
Thermo Scientific Maxima™ Reverse Transcriptase products are designed for consistency and efficiency in cDNA synthesis for RT-PCR and RT-qPCR applications. (thermofisher.com)
With these features, Maxima H Minus Reverse Transcriptase offers a robust performance in first strand cDNA synthesis. (thermofisher.com)
In RT-qPCR applications, Maxima H Minus Reverse Transcriptase also enables high efficiency synthesis over a wide range of input template amounts providing sensitive and accurate quantification of cDNA. (thermofisher.com)
Maxima H Minus Reverse Transcriptase efficiently synthesizes cDNA from a wide range of template amounts, has higher yields, and better linearity in cDNA synthesis outperforming other commercially available reverse transcriptases, making it an exceptional choice for RT-qPCR experiments (Figure 3) . (thermofisher.com)
The premixed solutions in the Thermo Scientific Maxima First Strand cDNA Synthesis Kits help further improve reproducibility and reaction setup time. (thermofisher.com)
Maxima H Minus First Strand cDNA Synthesis kit demonstrates consistently better reverse transcription efficiency than competitor kits. (thermofisher.com)
Maxima Reverse Transcriptases and Maxima H Minus Reverse Transcriptases are available in a variety of formats including stand-alone enzymes, kits, and master mixes. (thermofisher.com)
Maxima H Minus Reverse Transcriptase is engineered with molecular evolution techniques selecting for mutations that confer dramatically improved thermostability, processivity, and activity rates compared to wild-type M-MuLV enzymes. (thermofisher.com)
Maxima H Minus RT has 50X increased processivity compared with wild type MMuLV reverse transcriptase enzymes. (thermofisher.com)
Thermostability2
The engineered WarmStart Luna Reverse Transcriptase also possesses higher thermostability than many other RTs, allowing an optimal reaction temperature of 55°C. For difficult targets/templates, higher RT step temperatures of up to 60°C can be used without compromising Luna performance. (neb.com)
Maxima H Minus Reverse Transcriptase has diminished RNase H activity but retains all features of Maxima Reverse Transcriptase such as thermostability and enhanced processivity that have been improved. (thermofisher.com)
Viral2
Without reverse transcriptase, the viral genome would not be able to incorporate into the host cell, resulting in failure to replicate. (wikipedia.org)
Reverse transcriptase then adds DNAnucleotides onto the 3' end of the primer, synthesizing DNA complementary to the U5 (non-coding region) and R region (a direct repeat found at both ends of the RNA molecule) of the viral RNA. (wikipedia.org)
Telomerase reverse transcriptase that maintains the telomeres of eukaryotic chromosomes. (wikipedia.org)
Transcription4
citation needed] The process of reverse transcription, also called retrotranscription or retrotras, is extremely error-prone, and it is during this step that mutations may occur. (wikipedia.org)
Its tolerance to high reaction temperatures allows efficient transcription of RNA regions with extensive secondary structure and helps to improve primer specificity, thereby increasing overall yields. (thermofisher.com)
Total RNA (1 μg) from human cells (lanes 1 and 2) or mouse cells (lanes 3 and 4) were used in a reverse transcription reaction with Maxima H Minus Reverse Transcriptase according to manufacturer's protocol. (thermofisher.com)
Because RNA is fragile and difficult to study, a strand of complementary DNA (cDNA) is synthesized from RNA, using reverse transcriptase during the RT-PCR procedure. (britannica.com)
Primers1
The initial PCR reaction amplifies a region of the HCV genome (External primers). (cdc.gov)
Genome1
Reverse transcriptases are used by viruses such as HIV and hepatitis B to replicate their genomes, by retrotransposon mobile genetic elements to proliferate within the host genome, and by eukaryotic cells to extend the telomeres at the ends of their linear chromosomes. (wikipedia.org)
Activation1
This temperature-dependent activation helps to prevent undesirable non-specific priming and extension prior to thermocycling, providing added security for setting up reactions at room temperature. (neb.com)
Procedure1
Samples found to be negative for HCV RNA were extracted a second time by the same procedure with an additional incubation at 50 degrees Celsius for 45 minutes with 25 units of reverse transcriptase (Boehringer Mannnheim, Inndianapolis) and 10 units of RNAsin (Boehringer Mannnheim). (cdc.gov)
Products2
Reaction products were resolved by alkaline gel electrophoresis. (thermofisher.com)
In some PCR reactions multiple sequences have identical nucleotide sequences, for example if five PCR products have identical sequences a 5 will be found in this column. (cdc.gov)
Targets1
Reverse transcriptase (RT) PCR is PCR performed on RNA targets. (medscape.com)
Temperature1
Maxima H Minus Reverse Transcriptase outperforms other reverse transcriptases in terms of producing high yields of full-length cDNA over a wide temperature range (Figure 1). (thermofisher.com)
Include1
Well-studied reverse transcriptases include: HIV-1 reverse transcriptase from human immunodeficiency virus type 1 (PDB: 1HMV) has two subunits, which have respective molecular weights of 66 and 51 kDas. (wikipedia.org)
Template2
Reverse-transcribing DNA viruses, such as the hepadnaviruses, can allow RNA to serve as a template in assembling and making DNA strands. (wikipedia.org)
citation needed] Reverse transcriptase creates double-stranded DNA from an RNA template. (wikipedia.org)
Discovery1
The discovery of this reaction earned Kary Mullis, a US scientist, the Nobel Prize for chemistry in 1993. (island.lk)
Virus1
RÉSUMÉ Peu de temps après les premières détections de cas d'infection par le virus de la grippe pandémique (H1N1) au Mexique, un nouveau système de surveillance du virus H1N1 a été mis en place dans l'Émirat d'Abu Dhabi en mai 2009. (who.int)
Full-length1
This reverse transcriptase is capable of synthesizing full-length cDNA from a wide range of RNA templates (Figure 2) . (thermofisher.com)
Method1
Therefore, we studied the relationship between the orexin and histamine systems of the CNS using electrophysiology, immunocytochemistry, and the reverse transcriptase (RT)-PCR method. (jneurosci.org)
Features1
The Reaction Mix features dUTP for carryover prevention and a non-fluorescent visible dye for monitoring reaction setup. (neb.com)
Laboratory2
The ability of reverse transcriptase to synthesize DNA from RNA has been used in the laboratory. (britannica.com)
This test, popularly known as the PCR, is a chemical reaction performed in a PCR machine under very strict laboratory conditions to avoid contaminations. (island.lk)
Mice2
Knock-in mice expressing a CCN1 mutant that is unable to bind α v β 5 /α v β 3 were impaired in ductular reaction, leading to massive hepatic necrosis and mortality after bile duct ligation (BDL), whereas treatment of these mice with soluble JAG1 rescued ductular reaction and reduced hepatic necrosis and mortality. (jci.org)
Blockade of integrin α v β 5 /α v β 3 , NF-κB, or NOTCH signaling in WT mice also resulted in defective ductular reaction after BDL. (jci.org)