30/08/2016 · DNA amplification tests are known to be highly sensitive and specific for the diagnosis of ocular chlamydial infection, and are superior to other laboratory methods such as tissue culture, antigen detection, and hybridisation. 2,3 More recently, newer nucleic acid amplification tests NAATs based on amplification of ribosomal RNA rRNA have. However, its clinical utility may be limited by lack of automation or reproducibility. Recent studies have suggested nucleic acid sequence-based amplification of target RNA may be more robust. In this study, nucleic acid sequence-based amplification was established to.
07/01/2002 · Comparison of the RNA-amplification based methods RT-PCR and NASBA for the detection of circulating tumour cells. Burchill SA1, Perebolte L, Johnston C, Top B, Selby P. Author information: 1Children's Cancer Research Laboratory, St. James's University Hospital, Leeds LS9 7TF, UK. s.a.burchill@leeds. Evidence has suggested that RNA may be a better target than DNA for detection of viable cells. The purpose of this study was to compare a real-time RNA amplification method nucleic acid sequence-based amplification or NASBA to a similar real-time DNA-based PCR method with respect to the ability to detect viable bacterial cells. Comparison of the RNA-amplification based methods RT–PCR and NASBA for the detection of circulating tumour cells. Recent studies have suggested nucleic acid sequence-based amplification of target RNA may be more robust. Comparison of the RNA-ampliﬁcation based methods RT–PCR and NASBA for the detection of circulating tumour cells SA Burchill,1, L Perebolte3, C Johnston2, B Top3 and P Selby2 1Children’s Cancer Research Laboratory, St. James’s University Hospital, Leeds LS9 7TF, UK; 2ICRF Cancer Medicine Research Unit, St. James’s University.
15/12/2016 · In the present study, two systems based on this novel amplification technology, the Ovation Biotin System RS and a prototype system designed for target preparation from subnanogram level total RNA samples picogram Ribo-SPIA prototype system, or pRS, were evaluated and compared with the T7-based one- and two-round amplification systems. 20/07/2009 · Modified T7 RNA amplification. Total RNA from the BT474 breast cancer cell line and StratRef was linearly amplified through two rounds of in vitro transcription IVT based on a modified T7 amplification. Reverse transcription RT of total RNA was performed using 100 ng of oligo dT-T7 primer sequence provided in table 1 First. 31/10/2007 · On targeted arrays, we evaluated three amplification protocols, which are less time consuming than the commonly used T7-RNA polymerase based in vitro transcription protocols and therefore may be more suitable for clinical use: Template-switching polymerase chain reaction PCR, Ribo-single primer isothermal amplification and a random primer. DNA-based nucleic acid amplification tests NAATs are currently used to evaluate the success of treatment programmes by measuring the prevalence of C trachomatis infection. Some believe that newer ribosomal RNA rRNA-based tests may be much more sensitive since bacterial rRNA is present in amounts up to 10 000 times that of genomic DNA. 29/01/2015 · Generation of cDNA Using RNA Amplification Kits. The same batch of RNA and ERCC control mix was shipped to 12 different laboratories on dry ice. For comparison studies between amplification kits, each participant was instructed to combine 1 μl total RNA 1 μg/μl with 2 μl of 1:100 dilution of ERCC spike-in mix 1 and 7 μl water.
18/07/2017 · The RNA-based method, while was superior for detection of live bacterial cells still identified a number of 16S rRNA targets in samples spiked with dead cells. In environmental water samples, the DNA- and PMA-based approaches perhaps overestimated the richness of microbial community compared to RNA-based method. 03/06/2019 · Although developments in small RNA-Seq technology have improved detection of plasma-based miRNAs, the low RNA content and sequencing bias. This suggests that there may have been more miRNAs in the miRNeasy-extracted input RNA. Assuming amplification was within the exponential phase. Comparison of RNA extraction and.
19/02/2013 · Amplification of RNA-induced signals rather than the RNA sequence itself is a better choice to deal with the problem 10,20. A new isothermal reaction to simultaneously amplify and detect RNA based on the cleavage by DNAzyme and signal amplification is reported herein. Comparison of RNA extraction kits for the purification and. Each reaction had a total final volume of 25 μl which contained 5 μl of extracted sample RNA template. The reaction mix is based on the Qiagen. This amplification inhibition was resolved with pre-extraction homogenization or 1:10 dilution of the RNA prior to amplification. Comparison of an rRNA-based and DNA-based nucleic acid amplification test for the detection of Chlamydia trachomatis in trachoma Jon L Yang, Julius Schachter, Jeanne Moncada, Dereje Habte, Mulat Zerihun, Jenafir I House, Zhaoxia Zhou, Kevin C Hong, Kathryn Maxey, Bruce D. While these methods are highly promising, the amplification of the RNA sample itself is in more common use. RNA amplification is typically based on either T7 amplification of RNA, or PCR methods,,,. The advantage and disadvantage of these amplification methods have been described. 01/12/1997 · Comparison of the Amplicor HIV-1 monitor test and the nucleic acid sequence-based amplification assay for quantitation of human immunodeficiency virus RNA in plasma, serum, and plasma subjected to freeze-thaw cycles.
sequencing process. In comparison, 16S rRNA sequencing RNA-based can target live microbial cells in water as both dormant and metabolically active cells produce rRNA. The objective of this study was to compare the efficiency and sensitivity of DNA-based, PMA-based and RNA-based 16S rRNA Illumina. Samples Requirements. Targeted amplification-based NGS can be performed on a variety of samples, such as fresh or snap-frozen tissue, FFPE tissue, fine needle aspiration FNA samples, blood, bone marrow, buccal swabs, and cell pellets. Fresh and snap-frozen specimens are considered sources of the highest quality DNA and RNA with respect to. Recombinase polymerase amplification RPA is a single tube, isothermal alternative to the polymerase chain reaction PCR. By adding a reverse transcriptase enzyme to an RPA reaction it can detect RNA as well as DNA, without the need for a separate step to produce cDNA.
|07/01/2002 · Nucleic acid sequence-based amplification. Nucleic acid sequence-based amplification assays were established using cell line RNA at the laboratories of Organon Teknika Boxtel, The Netherlands. Briefly, multiple primer and probe sets for each of the mRNA targets were developed and tested in serial dilutions of cell line RNA.||25/01/2002 · Nucleic acid sequence-based amplification. Nucleic acid sequence-based amplification assays were established using cell line RNA at the laboratories of Organon Teknika Boxtel, The Netherlands. Briefly, multiple primer and probe sets for each of the mRNA targets were developed and tested in serial dilutions of cell line RNA.|
For microarray experiments starting with nanogram amounts of RNA it is essential to implement reproducible and powerful RNA amplification techniques. Available methods were mainly tested for reproducibility, only a few studies concentrated on potential amplification bias. We evaluated three amplification protocols, which are less time-consuming. Transcription-mediated amplification TMA is an isothermal does not change the nucleic acid temperature, single-tube nucleic acid amplification system utilizing two enzymes, RNA polymerase and reverse transcriptase.
01/11/2003 · Comparison of the NucliSens Basic Kit Nucleic Acid Sequence-Based Amplification and the Argene Biosoft Enterovirus Consensus Reverse Transcription-PCR Assays for Rapid Detection of Enterovirus RNA in Clinical Specimens. Similar to the PCR-based amplification of HCV RNA from liver biopsy specimen, 5 μL of extracted RNA from each dilution step was transferred directly to the TMA amplification step. For amplification of target RNA, two enzymes reverse transcriptase RT and RNA polymerase are used. About Cookies, including instructions on how to turn off cookies if you wish to do so. By continuing to browse this site you agree to us using cookies as described in About Cookies.
30/07/2015 · RNA-Seq has enabled high-throughput gene expression profiling to provide insight into the functional link between genotype and phenotype. Low quantities of starting RNA can be a severe hindrance for studies that aim to utilize RNA-Seq. To mitigate this bottleneck, whole transcriptome amplification WTA technologies have been developed to. A Comparison of Methods for DNA and RNA Purification from Plant. performed well in amplification-based applications. RNA Yield and Purity RNA yield was obtained from each plant leaf tissue using the two methods is shown in Figure 2, Panel A. We examined the relative efficiency and specificity of the real-time nucleic acid sequence-based amplification NASBA comparing with the established reverse transcription polymerase chain reaction RT-PCR and viral culture method which were used for the detection of enterovirus RNA. 17/12/2014 · Comparison of three RNA-Amplification protocols at the single cell level. As the aim of our study was to identify a flexible, sensitive and reproducible protocol that could be used to transcriptionally profile at the single cell level initial experiments aimed to directly compare cDNA generated using three kits that were commercially available.
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