Fingerprint Dive into the research topics of Feline coronavirus quantitative reverse-transcriptase polymerase chain reaction on effusion samples in cats with and without feline infectious peritonitis. Together they form a unique fingerprint. ...
Computed tomography (CT) of the chest is commonly used in the clinical management and assessment of complications of pneumonia caused by the novel coronavirus-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-, as well as to exclude alternative diagnoses(1). Although chest CT plays an important role, it should not be utilized as a screening tool and it can not be used by itself to confirm or exclude the diagnosis of this entity known as COVID-19(2,3), first described in the city of Wuhan, in the province of Hubei, in China, and declared as a pandemic by the World Health Organization in March 2020(4).. The diagnosis of COVID-19 is confirmed by real-time reverse-transcriptase polymerase chain reaction (RT-PCR) and partial or complete sequencing of the viral genome from nasopharyngeal aspirates, nasal and oral swabs, sputum or tracheal or bronchial lavage(5). The vast majority of patients with SARS-CoV-2 is asymptomatic, but the spectrum of clinical presentation is broad including ...
A quantitative real time reverse-transcription polymerase chain reaction (qRT-PCR) assay with specific primers recommended by the World Health Organization (WHO) has been widely used successfully for detection and monitoring of the pandemic H1N1/2009 influenza A virus. In this study, we report the design and characterization of a novel set of primers to be used in a qRT-PCR assay for detecting the pandemic H1N1/2009 virus. The newly designed primers target three regions that are highly conserved among the hemagglutinin (HA) genes of the pandemic H1N1/2009 viruses and are different from those targeted by the WHO-recommended primers. The qRT-PCR assays with the newly designed primers are highly specific, and as specific as the WHO-recommended primers for detecting pandemic H1N1/2009 viruses and other influenza viruses including influenza B viruses and influenza A viruses of human, swine, and raccoon dog origin. Furthermore, the qRT-PCR assays with the newly designed primers appeared to be at least 10-fold
TY - JOUR. T1 - Suppression subtractive hybridization. AU - Ghorbel, Mohamed T. AU - Murphy, David. PY - 2011. Y1 - 2011. N2 - Comparing two RNA populations that differ from the effects of a single independent variable, such as a drug treatment or a specific genetic defect, can establish differences in the abundance of specific transcripts that vary in a population dependent manner. There are different methods for identifying differentially expressed genes. These methods include microarray, Serial Analysis of Gene Expression (SAGE), and quantitative Reverse-Transcriptase Polymerase Chain Reaction (qRT-PCR). Herein, the protocol describes an easy and cost-effective alternative that does not require prior knowledge of the transcriptomes under examination. It is specifically relevant when low levels of RNA starting material are available. This protocol describes the use of Switching Mechanism At RNA Termini Polymerase Chain Reaction (SMART-PCR) to amplify cDNA from small amounts of RNA. The ...
The COVID-19 pandemic began in Wuhan (Hubei, China) in December 2019 and rapidly spread around the world. At the beginning of April 2020, there were over 100,000 confirmed cases and more than 10,000 deaths in Spain. Radiological literature has until now focused on non-contrast computed tomography (CT) findings to describe the probability of infection due to limitations of sensitive real-time reverse-transcription polymerase chain reaction (RT-PCR) results ...
TY - JOUR. T1 - Quantitative RT-PCR assay for HPV infection in cultured cells. AU - Culp, Timothy D.. AU - Christensen, Neil D.. PY - 2003/8/1. Y1 - 2003/8/1. N2 - Early events in the life cycle of the human papillomaviruses (HPV) have been difficult to investigate due to both the scarcity of authentic HPV virions and limitations in assays to detect and quantify nonpermissive infections in monolayer cell culture. We have developed a quantitative reverse transcription-PCR (QRT-PCR) assay for the E1 ^ E4 transcript of HPV-11. This assay is both sensitive, and capable of differentiating between infections caused by a wide range of virus input. The QRT-PCR assay measured accurately the relative amount of viral transcripts present in samples during validation experiments using RNAs from three cell lines. Infections in all three cell lines, using titrations of HPV-11 virions ranging from 20 to 600 particles per cell, produced linear expression profiles suggesting that these multiplicities of infection ...
TY - JOUR. T1 - Real-time quantitative reverse transcriptase polymerase chain reaction.. AU - Fan, Hongxin. AU - Robetorye, Ryan S.. PY - 2010. Y1 - 2010. N2 - The real-time quantitative reverse transcriptase polymerase chain reaction (RQ-PCR) has become the method of choice for the quantification of specific mRNAs. This method is fast, extremely sensitive, and accurate, requires only very small amounts of input RNA, and is relatively simple to perform. These characteristics have made it the method of choice for minimal residual disease monitoring such as in chronic myelogenous leukemia (CML). CML comprises approximately 20% of all leukemias and is characterized by a balanced (9;22) chromosomal translocation that results in the formation of a chimeric gene comprised of the BCR (breakpoint cluster region) gene and the ABL oncogene (BCR-ABL fusion gene). The chimeric gene encodes a fusion protein with constitutively increased tyrosine kinase activity, resulting in growth factor-independent ...
PubMedID: 23777485 | Quantitative expression analysis and prognostic significance of the BCL2-associated X gene in nasopharyngeal carcinoma: a retrospective cohort study. | BMC cancer | 1/1/2013
In final result, this analysis implies a surrounding function for coinfecting infections in clients introducing with SARI and highlights the crucial role of viral coinfection. Persisted use of the rRT-PCR multiplex assay in combination with the SARI surveillance program will boost our capability to find blood circulation of respiratory trojans in clients hospitalized for SARI and aid explain the factor of these respiratory system viruses among individuals with SARI in Southwest Photography equipment. Although there happen to be increasing problems about the prospective for A(H1N1)pdm09 to reassort with pre-existing real human influenza infections and give go up to a very transmissible or pathogenic pathogen [22], no blended infections have been noticed with either subtypes in individuals with SARI in this analysis.. A lymphocytic meningitis had been current in 6 of 21 HIV -seropositive people but in zero of the HIV -seronegative patients. Perivascular infiltrates consisting of lymphocytes and ...
The dysregulated circular RNAs (circRNAs) are relevant to the development of osteoarthritis (OA). The circRNA serpin family E member 2 (circSERPINE2) is dysregulated in OA, while the role and mechanism of circSERPINE2 in OA are largely unknown. The aim of our research is to explore how and whether circSERPINE2 regulates interleukin-1β (IL-1β)-caused chondrocyte damage in OA. In the present study, the chondrocytes (CHON-001 cells) were exposed to IL-1β to mimic the injury in OA. CircSERPINE2, microRNA-495 (miR-495) and transforming growth factor-β receptor 2 (TGFBR2) abundances were detected via quantitative reverse-transcription polymerase chain reaction (qRT-PCR) or Western blot. Cell apoptosis was assessed via viability, apoptotic rate and caspase-3 activity. Extracellular matrix was investigated by levels of Sry-type high-mobility-group box 9 (SOX9), collagen type II α 1 (COL2A1) and Aggrecan using Western blot. The interaction among circSERPINE2, miR-495 and TGFBR2 was assessed via ...
ONC201/TIC10 is a first-in-class small molecule inducer of TRAIL that causes early activation of the integrated stress response. Its promising safety profile and broad-spectrum efficacy in vitro have been confirmed in Phase I/II trials in several advanced malignancies. Binding and reporter assays have shown that ONC201 is a selective antagonist of the dopamine D2-like receptors, specifically, DRD2 and DRD3. We hypothesized that ONC201s interaction with DRD2 plays a role in ONC201s anticancer effects. Using cBioportal and quantitative reverse-transcription polymerase chain reaction analyses, we confirmed that DRD2 is expressed in different cancer cell types in a cell type-specific manner ...
The 2014 NCCN Clinical Practice Guidelines in Oncology for Chronic Myelogenous Leukemia recommend quantitative reverse-transcription polymerase chain reaction (QPCR) standardized to International Scale (IS) as the preferred method for monitoring molecular response to tyrosine kinase inhibitor (TKI) therapy. A BCR-ABL1 transcript level of 10% or less (IS) is now included as the response milestone at 3 and 6 months. Change of therapy to an alternate TKI is recommended for patients with BCR-ABL1 transcript levels greater than 10% (IS) at 3 months after primary treatment with imatinib. Continuing the same dose of TKI or switching to an alternate TKI are options for patients with BCR-ABL1 transcript levels greater than 10% (IS) at 3 months after primary treatment with dasatinib or nilotinib. The guidelines recommend 6-month evaluation with QPCR (IS) for patients with BCR-ABL1 transcript levels greater than 10% at 3 months. Monitoring with QPCR (IS) every 3 months is recommended for all patients, ...
Rani S, ODriscoll L., Reverse-transcriptase polymerase chain reaction to detect extracellular mRNAs., Methods Mol Biol., 784, 2011, 15 - 25 ...
Vela Diagnostics has launched the Zika Virus RT-PCR Test to be used on the firms Sentosa PCR workflow. The test can be used in conjunction with the companys other tropical infectious disease tests. Vela previously launched the CE-IVD Sentosa SA Chikungunya RT-PCR Test and the Sentosa Dengue I-IV RT-PCR Test as part of its Tropical Infections Panel.
The Deoxy+ OneStep RT-PCR Kit is a ready-to-use master mix which eliminates the need for optimization of reaction and cycling conditions for one-step RT-PCR. The reaction can be prepared by simply adding template RNA and primers to the master mix. The use of Yeasterns Hotstart DNA polymerase and Deoxy+ HiSpec RT enables reliable real-time RT-PCR quantification on any real-time PCR machines. Since it is a one-tube reaction, the procedure makes high-throughput analysis possible.. After reverse transcription, reactions are heated to 95°C for 10 minutes to inactivate the reverse transcriptase and simultaneously activate HotStart Taq DNA polymerase. This hot start to the PCR eliminates any nonspecific amplification products such as primer-dimers and reduces background smear, ensuring highly sensitive and reproducible RT-PCR.. ...
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How can one calculate the efficiency of isolated total RNA from different tissues treated at the same time, when it comes to doing quantitative real-time RT-PCR analysis afterwards? If one compare the real-time expression of a certain gene in different tissues, using housekeeping genes and an external standard reference gene.. How can one be sure that the possible difference in the expression pattern is not due to difference in the isolation efficiency?. ...
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subject: discussion on Quantitative Competitive RT-PCR dear netters, I am busy writing a discussion for a report I hope graduate on. So at the moment I am looking for some additional comments and insights on Quantitative Competitive RT-PCR that will help me getting a more thorough discussion. Here are some of the questions I am taking in account: What are the strong and weak points in the QC RT-PCR system (internal control: insertion of approx. 4bp; 5 fluorescent labeled primer; genescan analysis/ABI 370)? What is the best way to validate the method you are trying to set up. And are the results obtained with this method reliable/reproducible enough to get accepted by reviewers. Is there some strong literature discussing the system including the disadvantages of it. Thanks in advance, David E. Sluijs Pathology, Academic Hospital Utrecht. please mail to: ,paspoort at dds.nl ...
Reverse transcription-polymerase chain reaction: …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.
Nowadays the role of genetic findings in determining the diagnosis, therapy and prognosis of acute myeloid leukemia (AML) has become more valuable. To improve and validate the detection of clonal chromosomal aberrations in leukemia, we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML. Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients, and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH. The PCR products in some suspected cases were tested by two-directional sequencing. The results showed that except unqualified samples, fusion genes were detected by multiplex RT-PCR in 211 of 474 patients (44.51%), including AML1-ETO, CBFβ-MYH11, PML-RARα, ...
A reverse transcription-polymerase chain reaction (RT-PCR) test is the only way to identify an infected COVID-19 person. Since it takes 24 to 36 hours to achieve results, the Indian Council for Medical Research (ICMR) recently introduced Chinese-made rapid tests kits which can produce results in 10 to 15 minutes. However, this exercise didnt prove successful because these test kits showed erratic results.. A lot of patients have complained that their first RT-PCR test was negative but later on, when the COVID-19 symptoms persisted and when they went for the second test, it came positive. They blame the diagnostic labs for these inconsistencies.. Diagnostic labs, which ICMR has approved for COVID-19 tests, say that the RT-PCR test can only detect 65 out of 100 positive patients because it has its own limitation. A whole gamut of factors decides the accuracy of the test.. This also explains why many people who have tested negative initially become positive later on.. Experts say that the presence ...
Mercedes Pérez-Ruiz, José-María Navarro-Marí, María-Fé Bautista-Marín, Irene Pedrosa-Corral, Sara Sanbonmatsu-Gámez, Ana G. Camacho, José Rojas, Jorge Ruiz-Ortiz, Javier Rodríguez-Granger, José Antonio Carrillo ...
GermOnline 4.0 is a portal that cross-species database which focuses on high-throughput expression data relevant to the development of the…. Read More ...
Some gene transcripts represented by more than one probe set (each detected similar levels of expression) Cancer Genome Anatoy Project verified expression of many genes Q-RT-PCR -- some of 100 genes selected and measured for transcript levels using Q-RT-PCR --results in agreement with relative levels of microarray gene expression Immunohisotchemical analysis of tumor samples: established protein expression levels of select 100 correlated with mRNA levels from microarray Several genes identified have already been previously shown to be differentially expressed in metastatic prostate cancer ...
SOLIScript 1-step Multiplex Probe Kit is optimized for probe based one-step real-time quantitative RT-PCR assays. Learn about the product and order free samples here!
Lets set the record straight about DDRT-PCR! This technique recieves an amazing amount of bad press, which I find utterly astonishing given that has major advantages over other methods for identifying differentially expressed genes. It is an extremely powerful tool. I work in a research group of 6 scientists and we all routinely use DDRT-PCR for a variety of different purposes, using RNA extracted from a wide variety of material, ranging from cultured cells, specific tissues to whole embryos. We have been using the technique for the last couple of years with a great deal of success. My particular project is concerned with hunting for brain transcripts up- or down-regulated in developing mouse brain as a direct result of the targeted deletion of a specific gene. This is designed to give us clues as to the function of the encoded protein (the null has no phenotype). For this project I am using whole mouse brains from a variety of wt/null developmental timepoints, which DDRT-PCR allows me to ...
What could be the possible reasons for a RT-PCR experiment that was working fine - posted in PCR, RT-PCR and Real-Time PCR: What could be the possible reasons for a RT-PCR experiment that was working fine to stop working, if nothing was changed? I seem to see quite a few questions about this. And Ive been battling with this problem for quite a few months now.... Thank you! Sakumi
Methods:This study used 63 peripheral blood and bone marrow (PB/BM) samples from children with ALL. Immunophenotyping of PB and BM samples were performed using flow cytometry to illustrate the lineage. Moreover, reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to amplify transcripts of leukemia-specific chromosome fusion gene ETV6/RUNX1 and to monitor the expression levels of the ETV6/RUNX1 in patients according to Van Dongen et al protocol ...
The first time we visit an establishm-ent or a health facility and a violation has been observed, we immediatel-y issue a warning. If we would still have the same observatio-n during the follow-up monitoring, or the next time we visit them, we will already issue a Notice of Violation. MARIA BELENDA Q. AMBI DTI 11 Regional Director
Two commercial PRRSV ELISA packages (IDEXX and Bionote) were evaluated for his or her sensitivity and specificity using 476 PRRS-positive serum examples collected from 7 animal challenge tests and 1,000 PRRS-negative sera. with the Chonbuk Country wide University Institutional GS-9350 Pet Care and Make use of Committee (acceptance amount: 2012-0025). 40 swine farms which have preserved PRRS-negative status within the last year were verified to be detrimental by real-time invert transcription-polymerase chain response (RT-PCR) and IDEXX PRRS 3XR Ab ELISA and had been selected for the analysis. Information concerning the primers and probe for the real-time RT-PCR is really as follows: ahead primer: TGTCAGATTCAGGGAGRATAAGTTAC; probe: TTTTGCACCACMGCCAGCCC; and invert primer: ATCARGCGCACAGTRTGATGC. RT-PCR was carried out using the AgPath-IDTM One-Step RT-PCR Package (Ambion, Austin, TX, U.S.A.) inside a 25 response quantity using 5 of extracted design template. PCR amplification included (a) GS-9350 ...
TAPSEL-Alhamdulillah, tenyata hasil swab MRS (62), PDP asal Batangtoru, Kabupaten Tapanuli Selatan, Sumatera Utara, negatif.. Bupati Tapsel, Syahrul M Pasaribu, yang saat ini juga menjabat sebagai Ketua Gugus Tugas Percepatan Penanganan COVID-19 mengatakan, mengacu ke surat Dinas Kesehatan Provinsi Sumut nomor.443.33/726.29/Dinkes/VI/2020 tanggal 12 Juni 2020, ternyata hasil swab PDP asal Batang Toru, negatif.. Dijelaskannya, hasil reverse transcriptase polymerase chain reaction (RT-PCR) dari swab ke 3 ( pada 4 Juni 2020) negatif. Selanjutnya, d swab ke 4 ( pada 6 Juni 2020) juga hasilnya negatif.. Dengan demikian MRS sudah bisa dinyatakan sembuh, bahkan sudah berkumpul kembali ke tengah keluarga.. MRS dinyatakan positif pada pemeriksaan swab pertama sementara swab kedua negatif. Bahkan akibat dari itu 4 orang keluarganya dilakukan Rapid Test kedua dengan hasil Non Reaktif.. Alhamdulillah, kabar ini tentu menggembirakan kita semua mengingat MRS di usia-nya yang sudah 62 tahun merupakan usia ...
EurobioPlex SARS-CoV-2 is a CE marked test based on real-time reverse-transcription and amplification designed for qualitative determination of absence or…
Fig. 4 Tnnt temporal expression patterns in developing zebrafish embryos as detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Total RNA was isolated from different developmental stages, and primers specific for sMyHC, Tnnt1, Tnnt2, Tnnt3a, and Tnnt3b were used for RT-PCR. β-actin was used as an internal control. Numbers in the right panel indicate the molecular mass of RT-PCR products. ...
In this article explain the finer points of the delta-delta-CT method to calculate up-/down- regulations. The following text is a writeup of a course I gave at a local highschool.
Hepcidin, a peptide hormone that decreases intestinal iron absorption and macrophage iron release, is a potential drug target for patients with iron overload syndromes because its levels are inappropriately low in these individuals. Endogenous stimulants of Hepcidin transcription include bone morphogenic protein 6 (BMP6) and interleukin-6 (IL-6) by effects on mothers against decapentaplegic homolog (Smad)4 or signal transducer and activator of transcription (Stat)3, respectively. We conducted a small-scale chemical screen in zebrafish embryos to identify small molecules that modulate hepcidin expression. We found that treatment with the isoflavone, genistein, from 28-52 hours postfertilization in zebrafish embryos enhanced Hepcidin transcript levels, as assessed by whole-mount in situ hybridization and quantitative real-time reverse-transcriptase polymerase chain reaction. Genisteins stimulatory effect was conserved in human hepatocytes: Genistein treatment of HepG2 cells increased both ...
Aberrant DNA methylation of CpG islands of cancer-related genes is among the earliest and most frequent alterations in cancerogenesis and might be of value for either diagnosing cancer or evaluating recurrent disease. This mechanism usually leads to inactivation of tumour-suppressor genes. We have designed the current study to validate our previous microarray data and to identify novel hypermethylated gene promoters. The validation assay was performed in a different set of 8 patients with colorectal cancer (CRC) by means quantitative reverse-transcriptase polymerase chain reaction analysis. The differential RNA expression profiles of three CRC cell lines before and after 5-aza-2-deoxycytidine treatment were compared to identify the hypermethylated genes. The DNA methylation status of these genes was evaluated by means of bisulphite genomic sequencing and methylation-specific polymerase chain reaction (MSP) in the 3 cell lines and in tumour tissues from 30 patients with CRC. Data from our previous
Aberrant DNA methylation of CpG islands of cancer-related genes is among the earliest and most frequent alterations in cancerogenesis and might be of value for either diagnosing cancer or evaluating recurrent disease. This mechanism usually leads to inactivation of tumour-suppressor genes. We have designed the current study to validate our previous microarray data and to identify novel hypermethylated gene promoters. The validation assay was performed in a different set of 8 patients with colorectal cancer (CRC) by means quantitative reverse-transcriptase polymerase chain reaction analysis. The differential RNA expression profiles of three CRC cell lines before and after 5-aza-2-deoxycytidine treatment were compared to identify the hypermethylated genes. The DNA methylation status of these genes was evaluated by means of bisulphite genomic sequencing and methylation-specific polymerase chain reaction (MSP) in the 3 cell lines and in tumour tissues from 30 patients with CRC. Data from our previous
Cancer stem cells are associated with metastatic potential, treatment resistance, and poor patient prognosis. Distant recurrence remains the major cause of mortality in rectal cancer patients with preoperative chemoradiotherapy (CRT). We investigated the role of three stem cell markers (CD133, OCT4, and SOX2) in rectal cancer and evaluated the association between these gene levels and clinical outcome in rectal cancer patients with preoperative CRT. Thirty-three patients with rectal cancer underwent preoperative CRT. Total RNAs of rectal cancer cells before and after CRT were isolated. Residual cancer cells after CRT were obtained from formalin-fixed paraffin-embedded (FFPE) specimens using microdissection. The expression levels of three stem cell genes were measured using real-time reverse-transcription polymerase chain reaction (RT-PCR). The association between these gene levels and radiation was evaluated using colon cancer cell lines. Immunohistochemical staining of these markers after CRT was also
Among diabetes-susceptibility genes in NOD mice, only Idd-1 has been clearly assigned: Idd-1 could be a gene complex composed of class II major histocompatibility complex (MHC) genes, I-A beta and I-E. Employing restriction fragment length polymorphism (RFLP) analysis and nucleotide sequencing, we revealed that ILI and CTS mice, which are nondiabetic but are derived from the same Jcl-ICR mice as NOD mice, share the same class II MHC genes with NOD mice suggesting that both ILI and CTS mice also possess susceptible Idd-1 genotype. This was supported by a breeding study. To compare the usage of T cell receptor (TCR) V beta genes in NOD mice with that in ILI mice, we employed quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) which revealed that TCR V beta usages of these mice were indistinguishable. RT-PCR method also revealed that the V beta transcript of T cells infiltrating into pancreas of NOD mice was not restricted but was rather diverse. Since NOD and ILI mice share the same
Goodell, C.K., Zhang, J., Strait, E., Harmon, K., Patnayak, D., Otterson, T., Culhane, M., Christopher-Hennings, J., Clement, T., Leslie-Steen, P., Hesse, R., Anderson, J., Skarbek, K., Vincent, A., Kitikoon, P., Swenson, S., Jenkins-Moore, M., McGill, J., Rauh, R., Nelson, W., OConnell, C., Shah, R., Wang, C., Main, R., Zimmerman, J.J. 2016. Ring test evaluation of the detection of influenza A virus in swine oral fluids by real-time reverse-transcription polymerase chain reaction and virus isolation. Canadian Journal of Veterinary Research. 80(1):12-20 ...
TY - JOUR. T1 - Cytokine and growth factor expression in Pagets disease: analysis by reverse-transcription/polymerase chain reaction. AU - Ralston, S H. AU - Hoey, S A. AU - Gallacher, S J. AU - Adamson, B B. AU - Boyle, I T. AU - Gordon, Sharon Andrea. PY - 1994. Y1 - 1994. N2 - We investigated expression of several cytokines and growth factors in explants of Pagetic and non-Pagetic bone samples using the technique of reverse-transcription/polymerase chain reaction (RT/PCR). Transcripts for IL-1 alpha and IL-1 beta, TNF-alpha, TNF-beta, IL-6, basic fibroblast growth factor (bFGF), transforming growth factor beta (TGF-beta) and insulin-like growth factor-I (IGF-I) were found to a variable degree in both Pagetic and non-Pagetic bone samples, but there was no significant difference in the patterns of expression for these factors in Pagetic bone (n = 18) as compared with non-Pagetic bone (n = 51). There was furthermore, no significant difference in the patterns of expression for the various ...
Five potential reference genes for RT-qPCR application, namely histone H3, beta-actin, GAPDH, ubiquitin and 18S rRNA, were evaluated for normalization of gene expression in four selected tissues (liver, kidney, thyroid and abdominal fat). Tissues were derived from fattening pigs exposed to different amounts and type of dietary iodine. Two software applications (geNorm and NormFinder) were used to evaluate the stability of the potential reference genes. All studied genes displayed high expression stability but different stability patterns between the investigated tissues. The results suggest GAPDH and 18S rRNA as reference genes applicable in all tissues investigated. Beta-actin and histone H3 are suitable reference genes for all tissues investigated except fat. In contrast, ubiquitin should be excluded from use as a reference gene in the porcine tissues analyzed due to variations in expression levels, despite the good expression stability.
A multiplex reverse transcription polymerase chain reaction (RT-PCR) protocol was evaluated for the simultaneous detection of Potato leaf roll virus (PLRV), Potato virus X (PVX) and Potato Virus Y (PVY). The multiplex RT-PCR detection of viruses was equally efficient whether RNAs were extracted using commercial kits or a sodium sulphite based nucleic acid extraction procedure. cDNAs were prepared either using a common primer (oligo dT) or specific antisense primer followed by specific primer pairs for PCR. The multiplex RT-PCR separation of strains of PVY was accomplished by using a competitive multiplex RT-PCR (with one antisense and two sense primers). The multi virus or multistrain detection approaches described here have potential application to other crop-virus combinations ...
Identification of potential reference genes. Potential reference genes are identified by (A) geNorm and (B) NormFinder. Low M-value or variability represents th
We report on the identification and genomic characterization of DEIN, a novel gene that exhibits a characteristic expression pattern in primary neuroblastoma. With a combined RACE and RT-PCR approach, the full-length sequences of the main transcript variants of this gene were characterized. According to SAGE, Northern blot, and quantitative real-time RT-PCR, variants A and B represent the major transcripts of DEIN in primary neuroblastoma. These isoforms are strongly expressed in stage IVS tumors, whereas lower expression levels were observed in stage IV tumors. In contrast, expression of transcript variant C did not differ in these stages, as determined by quantitative real-time RT-PCR, and seemed to be expressed at lower levels in neuroblastoma because it could not be detected by Northern blot hybridization and Ct values of quantitative real-time RT-PCR analysis were high (data not shown). In addition, low numbers of SAGE tag ATTTGTTACA corresponding to transcript D suggested that this isoform ...
To inform an example calculation, we use publicly published data for analytic specificity of the Quest Diagnostics reverse transcription polymerase chain reaction assay (likely 100%, minimum 95%, maximum 100%) and an informed but pessimistic assumption regarding the clinical sensitivity of the reverse transcription polymerase chain reaction assay (likely 90%, minimum 65%, maximum 99%). Estimation of population prevalence is challenging: the minimum in this scenario is based on a recent measurement of the prevalence of reverse transcription polymerase chain reaction positivity among asymptomatic individuals in Iceland (0.6%), while our maximum is based on a recently published estimate among asymptomatic parturients at a major academic center (13.8%).6,7 As is the case with nearly all measurements of disease prevalence, both of these estimates were measured in unique populations at specific points in time. We chose a most likely prevalence estimate of 1.0% based on preliminary, unpublished data ...
צעד אחד RT-PCR assay לגילוי וזיהוי genogroup של מבודד norovirus מ כיסאות ילדים, אשר מנצל primers ו בדיקות TaqMan ספציפיים מסגרת קריאה...
Reverse transcription polymerase chain reaction (RT‐PCR) expression patterns of the genes coding for Laccaria bicolor Zn finger protein and Ser/Threo protein
Quiescent and proliferating leukemia cells accumulate highly lethal DNA double-strand breaks that are repaired by 2 major mechanisms: BRCA-dependent homologous recombination and DNA-dependent protein kinase-mediated (DNA-PK-mediated) nonhomologous end-joining, whereas DNA repair pathways mediated by poly(ADP)ribose polymerase 1 (PARP1) serve as backups. Here we have designed a personalized medicine approach called gene expression and mutation analysis (GEMA) to identify BRCA- and DNA-PK-deficient leukemias either directly, using reverse transcription-quantitative PCR, microarrays, and flow cytometry, or indirectly, by the presence of oncogenes such as BCR-ABL1. DNA-PK-deficient quiescent leukemia cells and BRCA/DNA-PK-deficient proliferating leukemia cells were sensitive to PARP1 inhibitors that were administered alone or in combination with current antileukemic drugs. In conclusion, GEMA-guided targeting of PARP1 resulted in dual cellular synthetic lethality in quiescent and proliferating ...
Tissue RNA PrepMate™ uses a modified guanidium salt-lysis method to optimally extract total RNA from tissue. The amount of extracted total RNA can differ according to the tissue or cell type, so the starting amount should be adjusted to your needs. The extracted viral RNA can be used as a template of the RT-PCR or quantitative real time RT-PCR and cDNA synthesis, cDNA library construction, Microarray ...
Pregnancy in mammals featuring hemochorial placentation introduces a major conflict with the mothers immune system, which is dedicated to repelling invaders bearing foreign DNA and RNA. Numerous and highly sophisticated ...
This is an historical archive of the activities of the MRC Anatomical Neuropharmacology Unit (MRC ANU) that operated at the University of Oxford from 1985 until March 2015. The MRC ANU established a reputation for world-leading research on the brain, for training new generations of scientists, and for engaging the general public in neuroscience. The successes of the MRC ANU are now built upon at the MRC Brain Network Dynamics Unit at the University of Oxford.. ...
Naina Mishra. Tribune News Service. Chandigarh, July 29. The UT Administration is set to introduce the rapid antigen test for the first time for the diagnosis of Covid-19.. According to sources, the UT will procure around 2,000 kits in the coming days. A tender has been floated on the GeM portal.. The UT will use the antigen test for asymptomatic patients only. Those who test positive will be considered Covid patients. As per the ICMR advisory, symptomatic patients with negative antigen results have to go in for the RT-PCR test for confirmation. However, no RT-PCR test confirmation is required for asymptomatic individuals testing negative in the antigen test.. However, the antigen test has been widely condemned for its low sensitivity as compared to the RT-PCR test. In its advisory, the ICMR said it had evaluated the antigen kits performance in two labs and had found its sensitivity to be 50.6 per cent and 84 per cent.. Recently, the Delhi High Court had asked the Delhi Government why it was ...
Recently we have reported that intermediate-frequency magnetic field (IF-MF) exposure transiently altered the mRNA expression levels of memory function-related genes in the hippocampi of adult male mice. However, the effects of IF-MF exposure during brain development on neurological biomarkers have not yet been clarified. In the present study, we investigated the effect of IF-MF exposure during development on neurological and immunological markers in the mouse hippocampus in 3- and 7-week-old male mice. Pregnant C57BL/6J mice were exposed to IF-MF (21 kHz, 3.8 mT) for one hour per day from organogenesis period day 7 to 17. At adolescence, some IF-MF-exposed mice were further divided into exposure, recovery, and sham-exposure groups. The adolescent-exposure groups were exposed again to IF-MF from postnatal day 27 to 48. The expression of mRNA in the hippocampi was examined using a real-time RT-PCR method, and microglia activation was examined by immunohistochemical analysis. The expression levels of NR1
Using the laboratory RT-PCR method, we detect the presence of SARS-CoV-2 virus in the body. A negative result can be used to end the quarantine and is required for travelling to and from abroad. The test will take place in a general practitioner's office in Prague. You will usually find out the result the next day after collection. The result will be sent to you by e-mail.
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Molecular Cytogenetics and Multiplex Reverse-Transcriptase Polymerase Chain Reaction for Risk Stratification in Acute Myeloid Leukemia ...
Background: Current methodology of gene expression analysis limits the possibilities of comparison between cells/tissues of organs in which cell size and/or number changes as a consequence of the study (e.g. starvation). A method relating the abundance of specific mRNA copies per cell may allow direct comparison or different organs and/or changing physiological conditions. Methods: With a number of selected genes, we analysed the relationship of the number of bases and the fluorescence recorded at a present level using cDNA standards. A lineal relationship was found between the final number of bases and the length of the transcript. The constants of this equation and those of the relationship between fluorescence and number of bases in cDNA were determined and a general equation linking the length of the transcript and the initial number of copies of mRNA was deduced for a given pre-established fluorescence setting. This allowed the calculation of the concentration of the corresponding mRNAs per ...
There are currently no tests available over the counter or at a doctors office that can quickly detect and distinguish between the H7N9 virus and other flu viruses. However, a more sophisticated test that specifically detects H7N9 virus has been developed by CDC for use by qualified public health laboratories in the United States and internationally. This test involves collecting a respiratory tract (i.e., nose, throat, lung) sample from a sick patient. The sample is then sent to a public health laboratory where a procedure known as rRT-PCR (real-time reverse transcriptase polymerase chain reaction) is conducted. rRT-PCR is very accurate and sensitive at detecting flu viruses. This procedure typically provides results within 4 hours; however, the time involved in processing and reporting results may vary depending on the laboratory ...
Main focus of GENE QUANTIFICATION web page is to describe and summarize all technical aspects involved in quantitative gene expression analysis using real-time RT-PCR and competitive RT-PCR. It illustrates the usefulness of absolute and relative quantification assays in real-time PCR and real-time RT-PCR.
In order to identify biomarkers to predict the progression of Type 1 Diabetic Kidney Disease (DKD), we profiled the transcriptome (the set of RNA molecules produced) of leukocytes from 33 type 1 diabetic patients at the time of their enrollment in the study. Patients were followed for a minimum of five years and data were collected including information about the progression of DKD. We will identify genes that correlate to various outcomes with a focus on the decline of kidney function. To validate the expression, we will use a quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR,) a laboratory method that is used to quantify RNA. Validated genes will be further studied in the future with more patients.. ...
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ELISA and real-time qRT-PCR analyses demonstrated an increase of VEGF protein and mRNA levels in the inner ear after local EP2 or EP4 agonist application, indicating that the stimulation of EP2 and/or EP4 activates VEGF production in the inner ear ...
Cancer cells. (A) Relative expression levels of Nox1, 2, 3, 4, and 5 mRNAs in A549 cells were determined by real-time RT-PCR and are presented as mean delta Ct
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Test with Confidence. Test with ZEUS. Clinical Performance Studies Four separate cohorts of clinically characterized specimens were tested: COVID-19 RT-PCR Positive Patient Specimens (n=35). The date between EUA-approved PCR test result and specimen draw was 3 to 37 days, with an average of 15.97 days and a median of 14 days.COVID-19 RT-PCR Negative Patient Specimens…
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Supplementary Materials01. gene is certainly detected in mere 22% of one side inhabitants cells and in 78% of one non-side inhabitants cells. Whereas, AR gene appearance is within 100% one side inhabitants and non-side inhabitants cells isolated in the same individual prostate scientific specimen. These studies also show that executing RT-PCR on one cells isolated by FACS could be effectively executed to determine gene appearance in one cells from cell lines and enzymatically digested tissue. While these studies provide a simple yes/no expression readout, the more sensitive quantitative RT-PCR would be able to provide even more information if necessary. highly expressed in side populace cells [38] that can contribute to the side populace; KS-176 or (ii) though the single side populace cells possess functionally active ABCG2 transporter as evidenced by their ability to efflux DCV, the ABCG2 gene is not expressed in 100% side populace cells suggesting that the presence of a functionally active ...
Video created by Exploratorium for the course Tinkering Fundamentals: Motion and Mechanisms. A Chain Reaction machine is a deceptively simple concept, but one that allows for an incredibly complex and deep investigation into something we ...
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