extraction reverse transcription and quantitative real time pcr

... quantified by real- time quantitative reverse transcription RT -PCR using the iCycler iQ Real Time PCR (Bio-Rad) and QuantiTect SYBR PCR kits (Qiagen) Sense and antisense PCR primers were designed ... explants (a-c) Realexplants time RT -PCR assays demonstrating increased COX-2 gene expression after h and h in compressed explants versus control and no increase for COX-1 and COX-3 Standard curves ... compressive stress at h and h were analyzed by (a-c) real- time quantitative RT -PCR and (d) immunoblotting (a-c) Up-regulation of mPGES-1 mRNA expression but not cPGES and mPGES-2 mRNA expression...

Ngày tải lên: 09/08/2014, 08:22

... (Invitrogen) and diluted 1:100 before use for QPCR Quantitative PCR In order to measure housekeeping gene expression, cDNA was subject to QPCR using the platinum QPCR super mix kit (Invitrogen) and pre-designed ... dendritic cells and alters their immune function J Virol 2003, 77(8):4950-4959 Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real- time quantitative PCR and the 2(-Delta ... (Toledo and Towne strains) and VZV (Schenke strain) and harvested at the earliest time of maximal productive infection as illustrated in Table The expression levels of 10 commonly used QPCR normalisation...

Ngày tải lên: 18/06/2014, 18:20

... harvested at several time points (table 1) and RNA was extracted The RNA transcription level of putative reference genes was determined by quantitative real- time PCR as described below Extraction of ... design and coordination of the study and helped to draft the manuscript All authors read and approved the final manuscript Material and Methods Virus culture and virus detection by real- time PCR ... http://www.virologyj.com/content/2/1/7 The respective MOI and time of cell culture are shown in table and were chosen to allow maximal infection as determined by immunofluorescence and real- time PCR [8-11] For kinetic studies,...

Ngày tải lên: 18/06/2014, 22:20

... (Invitrogen) and diluted 1:100 before use for QPCR Quantitative PCR In order to measure housekeeping gene expression, cDNA was subject to QPCR using the platinum QPCR super mix kit (Invitrogen) and pre-designed ... dendritic cells and alters their immune function J Virol 2003, 77(8):4950-4959 Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real- time quantitative PCR and the 2(-Delta ... (Toledo and Towne strains) and VZV (Schenke strain) and harvested at the earliest time of maximal productive infection as illustrated in Table The expression levels of 10 commonly used QPCR normalisation...

Ngày tải lên: 20/06/2014, 01:20

... harvested at several time points (table 1) and RNA was extracted The RNA transcription level of putative reference genes was determined by quantitative real- time PCR as described below Extraction of ... design and coordination of the study and helped to draft the manuscript All authors read and approved the final manuscript Material and Methods Virus culture and virus detection by real- time PCR ... http://www.virologyj.com/content/2/1/7 The respective MOI and time of cell culture are shown in table and were chosen to allow maximal infection as determined by immunofluorescence and real- time PCR [8-11] For kinetic studies,...

Ngày tải lên: 20/06/2014, 04:20

... were tested for specificity by RTPCR and also by qRT -PCR, followed by a dissociation curve and agarose gel electrophoresis Real- time PCR conditions and analysis PCR reactions were performed in ... ubiquitin and elongation factor-1 alpha For species with both sexual and apomictic reproductive mode, the best reference genes for qRT -PCR experiments have not been reported yet Real time PCR has ... embryo sacs [9] Quantitative real- time PCR (qRT -PCR) has emerged as an important technique to compare the expression profiles of target genes in different species, tissues or treatments and also to...

Ngày tải lên: 12/08/2014, 03:20

... 0.4 μL of each primer and 1.25 U enzyme Real- time PCR Mitchell Real- time PCR was performed in an Mx 3005P QPCR system (Stratagene, La Jolla, CA) using a SYBR Green based PCR assay (with ROX as ... RPS13 CYP RPS13 SAND TUB SAND TUB EF1alpha SAND GAPDH RPS13 UBQ TUB UBQ UBQ EF1alpha TUB GAPDH TUB RAN1 RAN1 ACT SAND ACT GAPDH ACT SAND ACT SAND RAN1 CYP GAPDH TUB GAPDH GAPDH GAPDH SAND GAPDH GAPDH ... JW and MEC designed the experiments IM and SL performed the experiments IM performed data analysis and table and figure drawing MEC wrote the first draft, and IM, BH, SL, JW and MEC corrected and...

Ngày tải lên: 12/08/2014, 03:21

... fluorescent quantitative real- time PCR (FQ -PCR) This technology accurately quantifies target DNA in a given sample and then could accurately detect viral loads in clinical samples[34] Yang and Guo ... TaqMan real- time PCR for detection of hepatopancreatic parvovirus from Australia J Virol Methods 2007, 140:10-16 42 Hussein IT, Field HJ: Development of a quantitative real- time TaqMan PCR assay ... Arden KE, Nitsche A: Real- time PCR in virology Nucleic Acids Res 2002, 30:1292-1305 35 Yang F, Jia W, Yue H, Luo W, Chen X, Xie Y, Zen W, Yang W: Development of quantitative real- time polymerase chain...

Ngày tải lên: 12/08/2014, 04:21

... Experiments (cell culture and DNA microarray analysis) YD: Experiments (cell culture and quantitative real- time PCR analysis) YF: Experiments (cell culture and quantitative real- time PCR analysis) MI: ... cancer patients and lung cancer cells [1-3,6-10], marker gene candidates have varied depending on the report Quantitative real- time PCR (qPCR) is generally considered the “gold-standard” assay ... oligonucleotide array data via quantitative real- time PCR BMC Bioinformatics 2006, 7:23 Nardon E, Donada M, Bonin S, Dotti I, Stanta G: Higher random oligo concentration improves reverse transcription yield...

Ngày tải lên: 15/02/2014, 04:20

... quantitative real time PCR (qRT -PCR) has become widely accepted for rapid and reproducible quantification of gene expression Most previous attempts to quantify Vb expression using qRT -PCR used one ... slope and R value of standard curves for other reactions are listed in Table Specificity of the qRT -PCR qRT -PCR specificities were assessed by melting curve analysis and sequencing of amplified PCR ... and sequencing amplified PCR products Melting curve analysis and sequencing amplified PCR products of reactions for, Cb and Vb subgroups consisting of a single subgroup showed a single peak and...

Ngày tải lên: 18/06/2014, 16:20

Ngày tải lên: 14/08/2014, 07:21

... studies have given evidence for overall safety as well Quantitative real- time PCR (QRTPCR) is the most widely used method for specific quantitative assay of ultra low concentration of pDNA in ... http://www.gvt-journal.com/content/6/1/11 Conclusion Quantitative real- time PCR (QRTPCR) assay was developed to assess a residual pDNA vaccine pVAX-Hsp60 TM814 in mice and beef cattle In beef cattle, ultra ... volume of 20 μl using 2× Real time PCR Syber green master mix (Qiagen, Germany) with 0.5 μM primers: Page of 11 (page number not for citation purposes) Genetic Vaccines and Therapy 2008, 6:11 CMV-Hsp60-F:...

Ngày tải lên: 14/08/2014, 19:22

... reference gene for real- time reverse transcription PCR data normalization Anal Biochem 344: 141–143 17 23 Bustin SA, Mueller R (2005) Real- time reverse transcription PCR (qRT -PCR) and its potential ... Intercalating dye Quantitative real- time PCR (qPCR) [1] is a sensitive and robust technique directly evolved from the “end-point detection” polymerase chain reaction (PCR) [2] PCR is a polymerase-dependent ... characterized by slowdown trend of amplification and the products Roberto Biassoni and Alessandro Raso (eds.), Quantitative Real- Time PCR: Methods and Protocols, Methods in Molecular Biology, vol...

Ngày tải lên: 05/09/2015, 10:35

... Heid CA, Stevens J, Williams PM: Real time quantitative PCR Genome Res 1996, 6:986-994 36 Gibson UEM, Heid CA, Williams A: Novel method for real time quantitative RT -PCR Genome Res 1996, 6:995-1001 ... -2, and -3 in the synovium synthase-1, 2, and -3 and (OA) levels of the messages (RA) Relative expressionand rheumatoid arthritis for hyaluronanof knees in 2, and -3 and hyaluronidase-1, -2, and ... Society [32] SD, standard deviation Table Sequences of the gene-specific oligonucleotide primers and probes for real- time reverse transcriptase polymerase chain reaction Primer and probe HAS-1 Sequence...

Ngày tải lên: 09/08/2014, 01:24

... Analysis by Real- Time Quantitative PCR Quantitative PCR was performed using the Sequence Detector System, ABI PRISM 7500 (Applied Biosystems 7500 Fast Real- Time PCR System) PCR primers and the TaqMan ... QMZ carried out the real- time RT -PCR, participated in the clinical data collecting of the gastric carcinoma patients and drafted the manuscript LZH carried out the real- time RT -PCR ZZW participated ... cytometric methods [8] With the advent of quantitative real- time PCR techniques [9], precise quantification of a target sequence has become possible Quantitative PCR provides investigators not only...

Ngày tải lên: 18/06/2014, 16:20

... a PCR reaction containing approximately 100 copies of bacteriophage λ DNA and a primer pair directed against λ DNA The performance of the PCR was monitored by quantitative real- time PCR (qPCR) ... fluid of these 27 HSV PCR positive subjects The distribution is left censored and the majority of subjects had

Ngày tải lên: 19/06/2014, 08:20

... employing microtiter and transwell plates Semiquantitative polymerase chain reaction (PCR) and quantitative real- time PCR have been recently developed and employed for the detection and quantitation ... Figure A – C: Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral load A – C: Analysis of HA and quantitative real- time PCR data employed for the determination ... of real- time PCR was defined by calculating coefficients of variation of Ct values of replicates of standard curve dilutions [21,26] We first examined the fidelity of real- time quantitative PCR...

Ngày tải lên: 19/06/2014, 08:20

... a PCR reaction containing approximately 100 copies of bacteriophage λ DNA and a primer pair directed against λ DNA The performance of the PCR was monitored by quantitative real- time PCR (qPCR) ... fluid of these 27 HSV PCR positive subjects The distribution is left censored and the majority of subjects had

Ngày tải lên: 20/06/2014, 04:20

... employing microtiter and transwell plates Semiquantitative polymerase chain reaction (PCR) and quantitative real- time PCR have been recently developed and employed for the detection and quantitation ... Figure A – C: Analysis of HA and quantitative real- time PCR data employed for the determination of JC viral load A – C: Analysis of HA and quantitative real- time PCR data employed for the determination ... of real- time PCR was defined by calculating coefficients of variation of Ct values of replicates of standard curve dilutions [21,26] We first examined the fidelity of real- time quantitative PCR...

Ngày tải lên: 20/06/2014, 04:20

... Detection limit and standard curve of the multiplex real- time PCR The detection limit and standard curve of the multiplex real- time PCR was determined using S Typhimurium ATCC 14028 and S Enteritidis ... multiplex real- time PCR assay showed 100% specificity for Salmonella spp and S Typhimurium and 99.1% for S Enteritidis The multiplex real- time PCR assay showed 100% sensitivity for Salmonella spp and ... the results of the standard culture method of isolated field and reference strains with those of the multiplex real- time PCR Results Specificity of the multiplex real- time PCR A total of 128 bacterial...

Ngày tải lên: 07/08/2014, 23:22