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    Qiagen rnalater
    Development of a candidate marker for ccRCC. (A) Expression values of CA9 correctly classified 30 of 32 samples in our FFPE dataset. (B) Whisker plot of expression value distribution in our FFPE dataset for CA9 . (C) Scatterplot for the expression values of CA9 in our FFPE and in our <t>RNAlater</t> dataset. (D) CA9 expression values correctly classify 139 out of 144 samples in a microarray dataset of ccRCC (GSE53757). (E) Distribution of CA9 expression values for normal (NO) and ccRCC tumor samples (TU) in the GSE53757 dataset. (F) Stratification of the expression values of overexpressed CA9 into all four stages of ccRCC [ 14 ].
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    Images

    1) Product Images from "Transcriptome Sequencing (RNAseq) Enables Utilization of Formalin-Fixed, Paraffin-Embedded Biopsies with Clear Cell Renal Cell Carcinoma for Exploration of Disease Biology and Biomarker Development"

    Article Title: Transcriptome Sequencing (RNAseq) Enables Utilization of Formalin-Fixed, Paraffin-Embedded Biopsies with Clear Cell Renal Cell Carcinoma for Exploration of Disease Biology and Biomarker Development

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0149743

    Development of a candidate marker for ccRCC. (A) Expression values of CA9 correctly classified 30 of 32 samples in our FFPE dataset. (B) Whisker plot of expression value distribution in our FFPE dataset for CA9 . (C) Scatterplot for the expression values of CA9 in our FFPE and in our RNAlater dataset. (D) CA9 expression values correctly classify 139 out of 144 samples in a microarray dataset of ccRCC (GSE53757). (E) Distribution of CA9 expression values for normal (NO) and ccRCC tumor samples (TU) in the GSE53757 dataset. (F) Stratification of the expression values of overexpressed CA9 into all four stages of ccRCC [ 14 ].
    Figure Legend Snippet: Development of a candidate marker for ccRCC. (A) Expression values of CA9 correctly classified 30 of 32 samples in our FFPE dataset. (B) Whisker plot of expression value distribution in our FFPE dataset for CA9 . (C) Scatterplot for the expression values of CA9 in our FFPE and in our RNAlater dataset. (D) CA9 expression values correctly classify 139 out of 144 samples in a microarray dataset of ccRCC (GSE53757). (E) Distribution of CA9 expression values for normal (NO) and ccRCC tumor samples (TU) in the GSE53757 dataset. (F) Stratification of the expression values of overexpressed CA9 into all four stages of ccRCC [ 14 ].

    Techniques Used: Marker, Expressing, Formalin-fixed Paraffin-Embedded, Whisker Assay, Microarray

    Gene network. The most differentially affected network with the central role of TGFB1 in (A) FFPE samples and B) RNAlater data sets. Proteins with cancer involvement are marked with purple outline . Red fill indicates overrepresentation of the gene in ccRCC , green indicates under-representation . Color intensity reflects range of fold change .
    Figure Legend Snippet: Gene network. The most differentially affected network with the central role of TGFB1 in (A) FFPE samples and B) RNAlater data sets. Proteins with cancer involvement are marked with purple outline . Red fill indicates overrepresentation of the gene in ccRCC , green indicates under-representation . Color intensity reflects range of fold change .

    Techniques Used: Formalin-fixed Paraffin-Embedded

    Pathway signature of VEGF and NOTCH mediated EMT in ccRCC. Comparison of gene expression data from the FFPE and from the RNAlater ® dataset with published results [ 20 ] and between themselves. F = FFPE samples , R = RNAlater ® samples , Numbers = fold change of up-regulation (red) or down-regulation (blue) .
    Figure Legend Snippet: Pathway signature of VEGF and NOTCH mediated EMT in ccRCC. Comparison of gene expression data from the FFPE and from the RNAlater ® dataset with published results [ 20 ] and between themselves. F = FFPE samples , R = RNAlater ® samples , Numbers = fold change of up-regulation (red) or down-regulation (blue) .

    Techniques Used: Expressing, Formalin-fixed Paraffin-Embedded

    Multidimensional scaling (MDS) analysis of gene expression data. MDS analysis based on all commonly detected genes shows that samples segregate by diagnosis (A) and not by storage condition (B). Distances correspond to leading log-fold-changes between each pair of samples. MDS based on differentially expressed genes demonstrates less within-group variance compared to MDS with all detected genes in the RNAlater ® (C) and FFPE (D) datasets. NF : Normal , FFPE; NR : Normal , RNAlater ® ; TF : Tumor , FFPE; TR : Tumor , RNAlater ® . NO = Normal; TU = Tumor .
    Figure Legend Snippet: Multidimensional scaling (MDS) analysis of gene expression data. MDS analysis based on all commonly detected genes shows that samples segregate by diagnosis (A) and not by storage condition (B). Distances correspond to leading log-fold-changes between each pair of samples. MDS based on differentially expressed genes demonstrates less within-group variance compared to MDS with all detected genes in the RNAlater ® (C) and FFPE (D) datasets. NF : Normal , FFPE; NR : Normal , RNAlater ® ; TF : Tumor , FFPE; TR : Tumor , RNAlater ® . NO = Normal; TU = Tumor .

    Techniques Used: Expressing, Formalin-fixed Paraffin-Embedded

    Immunohistochemistry and mRNA plots. (A) Immunohistochemistry of UMOD, NTPX2 and CA9. Magnification x20 , scale bar 50 μm . (B) Respective mRNA abundance plots in the FFPE and in the RNAlater ® datasets.
    Figure Legend Snippet: Immunohistochemistry and mRNA plots. (A) Immunohistochemistry of UMOD, NTPX2 and CA9. Magnification x20 , scale bar 50 μm . (B) Respective mRNA abundance plots in the FFPE and in the RNAlater ® datasets.

    Techniques Used: Immunohistochemistry, Formalin-fixed Paraffin-Embedded

    2) Product Images from "Adaptation of Iron Homeostasis Pathways by a Pseudomonas aeruginosa Pyoverdine Mutant in the Cystic Fibrosis Lung"

    Article Title: Adaptation of Iron Homeostasis Pathways by a Pseudomonas aeruginosa Pyoverdine Mutant in the Cystic Fibrosis Lung

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.01491-14

    prrF and hemO are expressed by P. aeruginosa infecting CF and bronchiectasis patients. Sputum from CF and bronchiectasis (Bron) patients chronically infected with P. aeruginosa was collected and preserved with RNALater. RNA was subsequently isolated and analyzed for expression of hemO and bphO (A) or prrF and prrH (B), and relative values were normalized to clpX and oprL as described in Materials and Methods. Samples 014-3 and 014 (A) were isolated at separate times (approximately 3 months apart) from the same CF patient. Relative levels of expression of hemO and bphO in PAO1 were determined in vitro in King's B medium.
    Figure Legend Snippet: prrF and hemO are expressed by P. aeruginosa infecting CF and bronchiectasis patients. Sputum from CF and bronchiectasis (Bron) patients chronically infected with P. aeruginosa was collected and preserved with RNALater. RNA was subsequently isolated and analyzed for expression of hemO and bphO (A) or prrF and prrH (B), and relative values were normalized to clpX and oprL as described in Materials and Methods. Samples 014-3 and 014 (A) were isolated at separate times (approximately 3 months apart) from the same CF patient. Relative levels of expression of hemO and bphO in PAO1 were determined in vitro in King's B medium.

    Techniques Used: Infection, Isolation, Expressing, In Vitro

    3) Product Images from "Neoadjuvant Therapy in Rectal Cancer - Biobanking of Preoperative Tumor Biopsies"

    Article Title: Neoadjuvant Therapy in Rectal Cancer - Biobanking of Preoperative Tumor Biopsies

    Journal: Scientific Reports

    doi: 10.1038/srep35589

    Of 195 analyzed patients (matched to available clinical data sets) 127 patients (65.13%) (good patients) were included for further molecular analyses (tumor content > 50%, RIN > 5). RNAlater biopsy samples not fullfilling the criterions (tumor content
    Figure Legend Snippet: Of 195 analyzed patients (matched to available clinical data sets) 127 patients (65.13%) (good patients) were included for further molecular analyses (tumor content > 50%, RIN > 5). RNAlater biopsy samples not fullfilling the criterions (tumor content

    Techniques Used:

    4) Product Images from "Use of RNAlater in fluorescence-activated cell sorting (FACS) reduces the fluorescence from GFP but not from DsRed"

    Article Title: Use of RNAlater in fluorescence-activated cell sorting (FACS) reduces the fluorescence from GFP but not from DsRed

    Journal: BMC Research Notes

    doi: 10.1186/1756-0500-3-328

    Fluorescence of DsRed2 protein is not affected in RNAlater . (A B) FACS results. All data in FACS figures are restricted to single cells defined by forward and side light scatter; clumped and ruptured cells (debris) are not displayed in the figure. DsRed positive and negative COS-7 cells were mixed before flow cytometry sorting. (A) Cells in BSA: a population of DsRed2 positive cells is clearly distinguished from DsRed2 negative cells. The cells with intermediate fluorescence intensity between the positive and negative populations represent newly dividing cells, which are in the initial stages of DsRed expression. (B) Cells in RNAlater: fewer cells are shown here than in (A) because RNAlater has induced cell clumping, so fewer singlets are available to the sorter. RNAlater did not quench fluorescent signals from analyzed DsRed positive cells. (C D) Dissociated cells were observed under the fluorescence microscope (C) DsRed2 positive cells in BSA. (D) DsRed2 positive cells after addition of RNAlater. The intensity of the DsRed2 was stable in the presence of RNAlater. (C D) Exposure time: 30ms; scale bar: 50 microns.
    Figure Legend Snippet: Fluorescence of DsRed2 protein is not affected in RNAlater . (A B) FACS results. All data in FACS figures are restricted to single cells defined by forward and side light scatter; clumped and ruptured cells (debris) are not displayed in the figure. DsRed positive and negative COS-7 cells were mixed before flow cytometry sorting. (A) Cells in BSA: a population of DsRed2 positive cells is clearly distinguished from DsRed2 negative cells. The cells with intermediate fluorescence intensity between the positive and negative populations represent newly dividing cells, which are in the initial stages of DsRed expression. (B) Cells in RNAlater: fewer cells are shown here than in (A) because RNAlater has induced cell clumping, so fewer singlets are available to the sorter. RNAlater did not quench fluorescent signals from analyzed DsRed positive cells. (C D) Dissociated cells were observed under the fluorescence microscope (C) DsRed2 positive cells in BSA. (D) DsRed2 positive cells after addition of RNAlater. The intensity of the DsRed2 was stable in the presence of RNAlater. (C D) Exposure time: 30ms; scale bar: 50 microns.

    Techniques Used: Fluorescence, FACS, Flow Cytometry, Cytometry, Expressing, Microscopy

    RNAlater decreases fluorescence of YFP . (A B) FACS results. All data in FACS figures are restricted to cells defined by forward and side light scatter and further to singlet events. The YFP positive cells are shown enclosed by red lines and negative cells are shown enclosed by black lines. (A) Cells in BSA: a population of YFP positive cells is clearly distinguished from YFP negative cells. (B) Cells in RNAlater: the two populations are not discernable. (C D) dissociated cells were observed under the fluorescence microscope (C) YFP positive cells in BSA. (D) YFP positive cells one minute after addition of RNAlater. The intensity of the YFP decreased substantially in the presence of RNAlater. (C D) Exposure time: 200ms; scale bar: 50 microns.
    Figure Legend Snippet: RNAlater decreases fluorescence of YFP . (A B) FACS results. All data in FACS figures are restricted to cells defined by forward and side light scatter and further to singlet events. The YFP positive cells are shown enclosed by red lines and negative cells are shown enclosed by black lines. (A) Cells in BSA: a population of YFP positive cells is clearly distinguished from YFP negative cells. (B) Cells in RNAlater: the two populations are not discernable. (C D) dissociated cells were observed under the fluorescence microscope (C) YFP positive cells in BSA. (D) YFP positive cells one minute after addition of RNAlater. The intensity of the YFP decreased substantially in the presence of RNAlater. (C D) Exposure time: 200ms; scale bar: 50 microns.

    Techniques Used: Fluorescence, FACS, Microscopy

    Fluorescence of Cy2 is not affected in RNAlater . (A B) FACS results. All data in FACS figures are restricted to cells defined by forward and side light scatter and further to singlet events. Dissociated cells were fixed and immunostained. The immunostained cells were visualized with secondary antibody conjugated with Cy2. The Cy2 positive cells are shown enclosed by red lines and negative cells are shown enclosed by black lines. (A) Cells in BSA: a population of Cy2 positive cells is clearly distinguished from Cy2 negative cells. (B) Cells in RNAlater: the two populations are distinguishable as well.
    Figure Legend Snippet: Fluorescence of Cy2 is not affected in RNAlater . (A B) FACS results. All data in FACS figures are restricted to cells defined by forward and side light scatter and further to singlet events. Dissociated cells were fixed and immunostained. The immunostained cells were visualized with secondary antibody conjugated with Cy2. The Cy2 positive cells are shown enclosed by red lines and negative cells are shown enclosed by black lines. (A) Cells in BSA: a population of Cy2 positive cells is clearly distinguished from Cy2 negative cells. (B) Cells in RNAlater: the two populations are distinguishable as well.

    Techniques Used: Fluorescence, FACS

    5) Product Images from "Human β-defensin-2 production from S. cerevisiae using the repressible MET17 promoter"

    Article Title: Human β-defensin-2 production from S. cerevisiae using the repressible MET17 promoter

    Journal: Microbial Cell Factories

    doi: 10.1186/s12934-017-0627-7

    Transcript levels determined by real-time PCR. hBD2 was expressed from a 2 µm-based expression plasmid containing either the MET17 or PRB1 promoter upstream of the hBD2 gene. These plasmids were transformed into S. cerevisiae strains DYB7 or DB1. The transformed yeast were inoculated at OD 600 = 0.15 into BMMD SFC without or with 1000 µM methionine and grown for 5 days, while cell pellet samples were taken approximately every 24 h and stored in RNAlater (Invitrogen) before RNA isolation and subsequently cDNA preparation. Error bars indicate coefficient of variation (n = 6). The fold difference is relative to the culture without methionine at 24 h in each strain (marked in red ). a hBD2 mRNA produced from DYB7 with the MET17 promoter under non-repressing (0 µM methionine) and repressing conditions (1000 µM methionine). b hBD2 mRNA produced from DYB7 with the PRB1 promoter without methionine added to the media (0 µM) and with added methionine (1000 µM). c hBD2 mRNA produced from DB1 with the MET17 promoter under non repressing (0 µM methionine) and repressing conditions (1000 µM methionine). TAF10 was used as the endogenous control
    Figure Legend Snippet: Transcript levels determined by real-time PCR. hBD2 was expressed from a 2 µm-based expression plasmid containing either the MET17 or PRB1 promoter upstream of the hBD2 gene. These plasmids were transformed into S. cerevisiae strains DYB7 or DB1. The transformed yeast were inoculated at OD 600 = 0.15 into BMMD SFC without or with 1000 µM methionine and grown for 5 days, while cell pellet samples were taken approximately every 24 h and stored in RNAlater (Invitrogen) before RNA isolation and subsequently cDNA preparation. Error bars indicate coefficient of variation (n = 6). The fold difference is relative to the culture without methionine at 24 h in each strain (marked in red ). a hBD2 mRNA produced from DYB7 with the MET17 promoter under non-repressing (0 µM methionine) and repressing conditions (1000 µM methionine). b hBD2 mRNA produced from DYB7 with the PRB1 promoter without methionine added to the media (0 µM) and with added methionine (1000 µM). c hBD2 mRNA produced from DB1 with the MET17 promoter under non repressing (0 µM methionine) and repressing conditions (1000 µM methionine). TAF10 was used as the endogenous control

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Plasmid Preparation, Transformation Assay, Isolation, Produced

    6) Product Images from "Inhalation of Ortho-Phthalaldehyde Vapor Causes Respiratory Sensitization in Mice"

    Article Title: Inhalation of Ortho-Phthalaldehyde Vapor Causes Respiratory Sensitization in Mice

    Journal: Journal of Allergy

    doi: 10.1155/2011/751052

    Effect of respiratory exposure to OPA vapor on the expression of Th2, Th1, and pro/anti-inflammatory cytokines in the lungs of mice. Mice were exposed to OPA (125, 250, 500, 1000 ppb) or filtered air according to the schedule shown in Figure 1 . Two days following the final exposure, the lungs were removed, inflated with RNALater, and processed for gene expression analysis. Data are presented as mean ( n = 5) and represent fold change relative to the concurrent control group. *Indicates that the cytokine was significantly increased at one or more of the OPA concentrations. Refer to Table S1 for the empirical gene expression data.
    Figure Legend Snippet: Effect of respiratory exposure to OPA vapor on the expression of Th2, Th1, and pro/anti-inflammatory cytokines in the lungs of mice. Mice were exposed to OPA (125, 250, 500, 1000 ppb) or filtered air according to the schedule shown in Figure 1 . Two days following the final exposure, the lungs were removed, inflated with RNALater, and processed for gene expression analysis. Data are presented as mean ( n = 5) and represent fold change relative to the concurrent control group. *Indicates that the cytokine was significantly increased at one or more of the OPA concentrations. Refer to Table S1 for the empirical gene expression data.

    Techniques Used: Expressing, Mouse Assay

    7) Product Images from "Cell differentiation defines acute and chronic infection cell types in Staphylococcus aureus"

    Article Title: Cell differentiation defines acute and chronic infection cell types in Staphylococcus aureus

    Journal: eLife

    doi: 10.7554/eLife.28023

    Experimental workflow to sort BRcells and DRcells using F luorescence A ctivated C ell S orting (FACS) to analyze and compare their transcriptomic profile. ( A ) Schematic flow of the experimental approach used for cell sorting and RNA-sequencing of specific cell types. Multicellular aggregates were resuspended and disaggregated in RNAlater for cell fixation. A homogenous suspension of single cells was obtained using mild sonication. FACS analysis separated the subpopulation of cells expressing the reporter in a test tube and the subpopulation of cells that did not express the reporter in a different test tube. We collected and concentrated the cells in a filter and isolated total RNA using hot phenol extraction protocol ( Blomberg et al., 1990 ). Total RNA was used to construct cDNA libraries that were sequenced using the Illumina HiSeq 2500. Results were analyzed using diverse bioinformatics tools. For further experimental procedures see Materials and Methods. ( B ) Control experiment of cell sorting. In the first panel, S. aureus wild type living cells from liquid cultures (non-fluorescent cells) were mixed with P ica -yfp labeled cells from liquid cultures (fluorescence cells) in relation 3:1 and analyzed using FACS. Second panel shows the flow cytometry analysis of sorted cells from the initial 3:1 mixture. The FACS-sorted bacterial population reached 97% enrichment of fluorescent cells. ( C ) Cell sorting of multicellular aggregates of S. aureus . A 4-days-old multicellular aggregate of P ica -yfp labeled strain was fixed using RNA later . Cells were dispersed and their fluorescence signal was analyzed using flow cytometry (left panel). Flow cytometry analysis showed higher expression of the reporter in a subpopulation of cells (subpopulation P1), as evidenced by the shoulder observed in the fluorescence expression profile of the culture. FACS of this sample led to an efficient separation of the subpopulation of fluorescence cells (P1) and the subpopulation of non-fluorescent cells (P2) in two different tubes. Flow cytometry analyses of P1 and P2 fluorescence signal showed that most of the cells from the P1 sample were fluorescent, whereas most of the cells from the P2 sample were non-fluorescent (center panel). In further experiments, we used a P ica -yfp labeled strain to separate a P1 subpopulation (BR+ sample) and a P2 subpopulation (BR- sample). Likewise, we used a P psmα -yfp labeled strain to separate a P1 subpopulation (DR+ sample) and a P2 subpopulation (DR- sample). We sorted approximately 25 million cells per sample prior RNA isolation. ( D ) Sorted fractions were diluted and plated on TSB and the resulting colonies were examined for viability and for emission of fluorescence using a fluorescence stereoscope. Consistent with our flow cytometry enrichment data, more than 95% of the colonies carried the transcriptional fusion and were fluorescent. In addition, the non-fluorescence cells (P2 fraction) revealed similar viability as the its fluorescent counterpart and. ( E ) PCR analysis of total RNA samples showed that samples are free of DNA contamination after DNaseI treatment. Amplification of rrna 16s control gene was detected only in the positive control (genomic DNA from S. aureus ). Molecular weight = 500 bp ladder. ( F ) Quantification of RNA concentration using spectrophotometry. The RNA concentration and absorption ratios for each sample were determined using the Nanodrop. Concentration is shown in the top right section of each panel. ( G ) Quality check of the RNA samples. RNA samples were examined using MultiNA microchip electrophoresis (Shimadzu) to determine quality and concentration of the RNA. After analysis, cDNA was synthesized following the protocol described in Materials and Methods. Molecular weight = 200 bp ladder. ( H ) Analysis of the PCR-amplified cDNA samples on a Shimadzu MultiNA microchip electrophoresis system. For Illumina sequencing, the cDNA was size-fractionated in the size range of 150–600 bp using a differential cleanup system. ( I ) An aliquot of each cDNA was analyzed by capillary electrophoresis. Each double-stranded cDNA sample was flanked with different adapter sequences to generate a cDNA with a combined length of 100 bases. Length range and concentration are shown at the top left section of each panel.
    Figure Legend Snippet: Experimental workflow to sort BRcells and DRcells using F luorescence A ctivated C ell S orting (FACS) to analyze and compare their transcriptomic profile. ( A ) Schematic flow of the experimental approach used for cell sorting and RNA-sequencing of specific cell types. Multicellular aggregates were resuspended and disaggregated in RNAlater for cell fixation. A homogenous suspension of single cells was obtained using mild sonication. FACS analysis separated the subpopulation of cells expressing the reporter in a test tube and the subpopulation of cells that did not express the reporter in a different test tube. We collected and concentrated the cells in a filter and isolated total RNA using hot phenol extraction protocol ( Blomberg et al., 1990 ). Total RNA was used to construct cDNA libraries that were sequenced using the Illumina HiSeq 2500. Results were analyzed using diverse bioinformatics tools. For further experimental procedures see Materials and Methods. ( B ) Control experiment of cell sorting. In the first panel, S. aureus wild type living cells from liquid cultures (non-fluorescent cells) were mixed with P ica -yfp labeled cells from liquid cultures (fluorescence cells) in relation 3:1 and analyzed using FACS. Second panel shows the flow cytometry analysis of sorted cells from the initial 3:1 mixture. The FACS-sorted bacterial population reached 97% enrichment of fluorescent cells. ( C ) Cell sorting of multicellular aggregates of S. aureus . A 4-days-old multicellular aggregate of P ica -yfp labeled strain was fixed using RNA later . Cells were dispersed and their fluorescence signal was analyzed using flow cytometry (left panel). Flow cytometry analysis showed higher expression of the reporter in a subpopulation of cells (subpopulation P1), as evidenced by the shoulder observed in the fluorescence expression profile of the culture. FACS of this sample led to an efficient separation of the subpopulation of fluorescence cells (P1) and the subpopulation of non-fluorescent cells (P2) in two different tubes. Flow cytometry analyses of P1 and P2 fluorescence signal showed that most of the cells from the P1 sample were fluorescent, whereas most of the cells from the P2 sample were non-fluorescent (center panel). In further experiments, we used a P ica -yfp labeled strain to separate a P1 subpopulation (BR+ sample) and a P2 subpopulation (BR- sample). Likewise, we used a P psmα -yfp labeled strain to separate a P1 subpopulation (DR+ sample) and a P2 subpopulation (DR- sample). We sorted approximately 25 million cells per sample prior RNA isolation. ( D ) Sorted fractions were diluted and plated on TSB and the resulting colonies were examined for viability and for emission of fluorescence using a fluorescence stereoscope. Consistent with our flow cytometry enrichment data, more than 95% of the colonies carried the transcriptional fusion and were fluorescent. In addition, the non-fluorescence cells (P2 fraction) revealed similar viability as the its fluorescent counterpart and. ( E ) PCR analysis of total RNA samples showed that samples are free of DNA contamination after DNaseI treatment. Amplification of rrna 16s control gene was detected only in the positive control (genomic DNA from S. aureus ). Molecular weight = 500 bp ladder. ( F ) Quantification of RNA concentration using spectrophotometry. The RNA concentration and absorption ratios for each sample were determined using the Nanodrop. Concentration is shown in the top right section of each panel. ( G ) Quality check of the RNA samples. RNA samples were examined using MultiNA microchip electrophoresis (Shimadzu) to determine quality and concentration of the RNA. After analysis, cDNA was synthesized following the protocol described in Materials and Methods. Molecular weight = 200 bp ladder. ( H ) Analysis of the PCR-amplified cDNA samples on a Shimadzu MultiNA microchip electrophoresis system. For Illumina sequencing, the cDNA was size-fractionated in the size range of 150–600 bp using a differential cleanup system. ( I ) An aliquot of each cDNA was analyzed by capillary electrophoresis. Each double-stranded cDNA sample was flanked with different adapter sequences to generate a cDNA with a combined length of 100 bases. Length range and concentration are shown at the top left section of each panel.

    Techniques Used: FACS, Flow Cytometry, RNA Sequencing Assay, Sonication, Expressing, Isolation, Construct, Labeling, Fluorescence, Cytometry, Polymerase Chain Reaction, Amplification, Positive Control, Molecular Weight, Concentration Assay, Spectrophotometry, MicroChIP Assay, Electrophoresis, Synthesized, Sequencing

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    BIA-KA:

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    Quantitative RT-PCR:

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    Expressing:

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    Article Snippet: The tissue was placed in RNAlater RNA stabilization reagent (Qiagen, Germantown, USA) and stored at 4°C. .. Real-time qRT-PCR for pea aphid tor was performed on a Step One Plus real-time thermocycler (Life Technologies, Grand Island, USA) using Taqman Gene Expression Master Mix (Life Technologies, Grand Island, USA) and a PrimeTime qPCR Assay (Integrated DNA Technologies, Coralville, USA) that spanned an intron-exon junction (probe 5'-/56-FAM/CAG CAT CAA /ZEN/ATC GTG GTA CGC CAA C/3IABkFQ/-3' with primers 5'-ATT CGA CTC CCT TGC TAT TCG-3' and 5'-TGG GTG ACT TGC AGA CTT AC-3').

    Article Title: Concomitant Interferon Alpha Stimulation and TLR3 Activation Induces Neuronal Expression of Depression-Related Genes That Are Elevated in the Brain of Suicidal Persons
    Article Snippet: Brain sample acquisition For gene expression analysis in the brain, samples were taken of each deceased individual as cubes with 3–5 mm edge length from different brain structures currently related to depression, such as hippocampus, amygdala and gyrus cinguli – . .. For RNA protection, the brain tissue was transferred immediately into 1 mL RNAlater RNA Stabilization Reagent (Qiagen) and stored over night at 4°C and then at −20°C.

    Genome Wide:

    Article Title: A Frameshift Mutation within LAMC2 Is Responsible for Herlitz Type Junctional Epidermolysis Bullosa (HJEB) in Black Headed Mutton Sheep
    Article Snippet: Genome-wide association analyses were performed using DNA from 12 HJEB-affected BHM lambs (cases), 6 HJEB-unaffected BHM sheep representing parents and full-sibs of the cases and 6 control samples from the whole BHM population. .. The samples were harvested immediately after euthanasia and asserved in RNAlater RNA Stabilization Reagent (Qiagen) for stabilization and protection of cellular RNA in situ and then stored at −20°C.

    Transplantation Assay:

    Article Title: MicroRNAs Induce Epigenetic Reprogramming and Suppress Malignant Phenotypes of Human Colon Cancer Cells
    Article Snippet: When tumor volumes reached 100 mm3 , as measured with calipers and calculated using the formula V = (ab2 )/2, where a is the length and b is the width, tumors were harvested and cut into 3–4 mm3 sections for serial transplantation. .. Xenograft tumors and mouse blood were collected during sacrifice and preserved in 10% neutral-buffered formalin for histology and immunohistochemical studies or in RNAlater RNA Stabilization Reagent (Qiagen, Tokyo, Japan).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: The pea aphid uses a version of the terminal system during oviparous, but not viviparous, development
    Article Snippet: The tissue was placed in RNAlater RNA stabilization reagent (Qiagen, Germantown, USA) and stored at 4°C. .. Real-time qRT-PCR for pea aphid tor was performed on a Step One Plus real-time thermocycler (Life Technologies, Grand Island, USA) using Taqman Gene Expression Master Mix (Life Technologies, Grand Island, USA) and a PrimeTime qPCR Assay (Integrated DNA Technologies, Coralville, USA) that spanned an intron-exon junction (probe 5'-/56-FAM/CAG CAT CAA /ZEN/ATC GTG GTA CGC CAA C/3IABkFQ/-3' with primers 5'-ATT CGA CTC CCT TGC TAT TCG-3' and 5'-TGG GTG ACT TGC AGA CTT AC-3').

    Immunohistochemistry:

    Article Title: MicroRNAs Induce Epigenetic Reprogramming and Suppress Malignant Phenotypes of Human Colon Cancer Cells
    Article Snippet: .. Xenograft tumors and mouse blood were collected during sacrifice and preserved in 10% neutral-buffered formalin for histology and immunohistochemical studies or in RNAlater RNA Stabilization Reagent (Qiagen, Tokyo, Japan). .. Carbonate apatite preparation To prepare a CA transfection mixture in vitro , 2 μg of each miR-302 (-a,-b,-c,-d), miR-369 (-3p, -5p) or NC miR was mixed with 4 μL of 1 M CaCl2 in 1 mL of serum-free bicarbonate (44 mM)-buffered DMEM medium (pH 7.5) incubated at 37°C for 30 min, and used for transfection [ – ].

    Article Title: Intravenous Inoculation of a Bat-Associated Rabies Virus Causes Lethal Encephalopathy in Mice through Invasion of the Brain via Neurosecretory Hypothalamic Fibers
    Article Snippet: Tissue harvest Brains and spinal cord for RNA isolation were immersed into an appropriate amount (1 ml per 100 mg tissue) of RNAlater RNA Stabilization reagent (Qiagen, www.qiagen.com/ ) immediately after harvest from the euthanized mouse and stored at 4°C for maximal four weeks until further processing. .. Brains and spinal cords determined for immunohistochemical analysis were immersion-fixed in Bouin Hollande fixative for 24 hours and washed with 70% isopropanol afterwards.

    Infection:

    Article Title: Infection- and procedure-dependent effects on pulmonary gene expression in the early phase of influenza A virus infection in mice
    Article Snippet: Mice were killed by CO2 asphyxiation at 6, 12, 18, 24, 48, and 120 h with respect to infection or mock treatment. .. Lungs were removed from the cadavers by cutting the main bronchus and were washed in RNAlater RNA Stabilization Reagent (Qiagen Inc, Valencia, CA, USA) immediately after removal.

    Article Title: The Absence of DHHC3 Affects Primary and Latent Herpes Simplex Virus 1 Infection
    Article Snippet: Paragraph title: Isolation of RNA from corneas and TG of infected mice. ... Isolated tissues were immersed in RNAlater RNA stabilization reagent (Qiagen, Germantown, MD) and stored at −80°C until they were processed.

    other:

    Article Title: Altered gene expression profiles in the hippocampus and prefrontal cortex of type 2 diabetic rats
    Article Snippet: Excised tissue samples were immediately fixed in RNAlater RNA stabilization reagent (Qiagen).

    Tumorigenicity Assay:

    Article Title: MicroRNAs Induce Epigenetic Reprogramming and Suppress Malignant Phenotypes of Human Colon Cancer Cells
    Article Snippet: The tumorigenicity assay was performed using an in vivo xenograft model. First, 5 × 106 cells suspended in a total volume of 200 μL DMEM/Matrigel (1:1 (v/v) suspension) were injected into the flanks of 7-8-week-old female mice (BALB/cAJcl-nu/nu). .. Xenograft tumors and mouse blood were collected during sacrifice and preserved in 10% neutral-buffered formalin for histology and immunohistochemical studies or in RNAlater RNA Stabilization Reagent (Qiagen, Tokyo, Japan).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: A Network Analysis of the Human T-Cell Activation Gene Network Identifies Jagged1 as a Therapeutic Target for Autoimmune Diseases
    Article Snippet: .. RNA extraction, probes and rtPCR Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque (Pharmacia Biotech) and they were immediately submerged in RNAlater RNA Stabilization Reagent (Qiagen) to preserve the gene expression patterns. .. Total RNA was isolated using the RNeasy Mini Kit (Qiagen), removing DNA with the RNase-Free DNase Set (Qiagen), and the High-Capacity cDNA Archive Kit (Applied Biosystems) was used to synthesise cDNA from the total RNA.

    Injection:

    Article Title: MicroRNAs Induce Epigenetic Reprogramming and Suppress Malignant Phenotypes of Human Colon Cancer Cells
    Article Snippet: The tumorigenicity assay was performed using an in vivo xenograft model. First, 5 × 106 cells suspended in a total volume of 200 μL DMEM/Matrigel (1:1 (v/v) suspension) were injected into the flanks of 7-8-week-old female mice (BALB/cAJcl-nu/nu). .. Xenograft tumors and mouse blood were collected during sacrifice and preserved in 10% neutral-buffered formalin for histology and immunohistochemical studies or in RNAlater RNA Stabilization Reagent (Qiagen, Tokyo, Japan).

    Cellular Antioxidant Activity Assay:

    Article Title: The pea aphid uses a version of the terminal system during oviparous, but not viviparous, development
    Article Snippet: The tissue was placed in RNAlater RNA stabilization reagent (Qiagen, Germantown, USA) and stored at 4°C. .. Real-time qRT-PCR for pea aphid tor was performed on a Step One Plus real-time thermocycler (Life Technologies, Grand Island, USA) using Taqman Gene Expression Master Mix (Life Technologies, Grand Island, USA) and a PrimeTime qPCR Assay (Integrated DNA Technologies, Coralville, USA) that spanned an intron-exon junction (probe 5'-/56-FAM/CAG CAT CAA /ZEN/ATC GTG GTA CGC CAA C/3IABkFQ/-3' with primers 5'-ATT CGA CTC CCT TGC TAT TCG-3' and 5'-TGG GTG ACT TGC AGA CTT AC-3').

    In Vivo:

    Article Title: MicroRNAs Induce Epigenetic Reprogramming and Suppress Malignant Phenotypes of Human Colon Cancer Cells
    Article Snippet: The tumorigenicity assay was performed using an in vivo xenograft model. First, 5 × 106 cells suspended in a total volume of 200 μL DMEM/Matrigel (1:1 (v/v) suspension) were injected into the flanks of 7-8-week-old female mice (BALB/cAJcl-nu/nu). .. Xenograft tumors and mouse blood were collected during sacrifice and preserved in 10% neutral-buffered formalin for histology and immunohistochemical studies or in RNAlater RNA Stabilization Reagent (Qiagen, Tokyo, Japan).

    Fluorescence:

    Article Title: A Network Analysis of the Human T-Cell Activation Gene Network Identifies Jagged1 as a Therapeutic Target for Autoimmune Diseases
    Article Snippet: RNA extraction, probes and rtPCR Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque (Pharmacia Biotech) and they were immediately submerged in RNAlater RNA Stabilization Reagent (Qiagen) to preserve the gene expression patterns. .. Primer sequences and target-specific fluorescence-labelled TaqMan probes were purchased from Applied Biosystems (TaqMan Gene Expression Assays, Supplementary ).

    Isolation:

    Article Title: The Absence of DHHC3 Affects Primary and Latent Herpes Simplex Virus 1 Infection
    Article Snippet: .. Isolated tissues were immersed in RNAlater RNA stabilization reagent (Qiagen, Germantown, MD) and stored at −80°C until they were processed. .. The corneas or TG from each animal were processed for RNA extraction using TRIzol reagent as we described previously ( ).

    Article Title: Platelet-derived β2M regulates monocyte inflammatory responses
    Article Snippet: Hearts from mice were collected day 0 or day 3 after MI and placed into RNAlater RNA Stabilization Reagent (Qiagen). .. Isolated hearts were homogenized using a Tissue-Tearor (BioSpec).

    Article Title: A Network Analysis of the Human T-Cell Activation Gene Network Identifies Jagged1 as a Therapeutic Target for Autoimmune Diseases
    Article Snippet: .. RNA extraction, probes and rtPCR Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque (Pharmacia Biotech) and they were immediately submerged in RNAlater RNA Stabilization Reagent (Qiagen) to preserve the gene expression patterns. .. Total RNA was isolated using the RNeasy Mini Kit (Qiagen), removing DNA with the RNase-Free DNase Set (Qiagen), and the High-Capacity cDNA Archive Kit (Applied Biosystems) was used to synthesise cDNA from the total RNA.

    Article Title: The pea aphid uses a version of the terminal system during oviparous, but not viviparous, development
    Article Snippet: The tissue was placed in RNAlater RNA stabilization reagent (Qiagen, Germantown, USA) and stored at 4°C. .. The RNA was purified using the SV Total RNA Isolation System (Promega, Madison, USA) and cDNA was made using the High Capacity cDNA Reverse Transcription Kit (Life Technologies, Grand Island, USA).

    Article Title: Intravenous Inoculation of a Bat-Associated Rabies Virus Causes Lethal Encephalopathy in Mice through Invasion of the Brain via Neurosecretory Hypothalamic Fibers
    Article Snippet: .. Tissue harvest Brains and spinal cord for RNA isolation were immersed into an appropriate amount (1 ml per 100 mg tissue) of RNAlater RNA Stabilization reagent (Qiagen, www.qiagen.com/ ) immediately after harvest from the euthanized mouse and stored at 4°C for maximal four weeks until further processing. .. Brains and spinal cords determined for immunohistochemical analysis were immersion-fixed in Bouin Hollande fixative for 24 hours and washed with 70% isopropanol afterwards.

    Article Title: Immune-Related Gene Expression Profile in Laboratory Common Marmosets Assessed by an Accurate Quantitative Real-Time PCR Using Selected Reference Genes
    Article Snippet: Paragraph title: RNA isolation ... Tissue samples (spleen, mesenteric lymph node, jejunum, ileum, descending colon, cerebrum, cerebellum, brainstem, heart, lung, liver and kidney) were excised from each animal and immediately submerged in RNAlater® RNA Stabilization Reagent (Qiagen).

    Article Title: Concomitant Interferon Alpha Stimulation and TLR3 Activation Induces Neuronal Expression of Depression-Related Genes That Are Elevated in the Brain of Suicidal Persons
    Article Snippet: For RNA protection, the brain tissue was transferred immediately into 1 mL RNAlater RNA Stabilization Reagent (Qiagen) and stored over night at 4°C and then at −20°C. .. Subsequently, total RNA was isolated according to the manufacturer's recommendations for lipid tissues using Qiazol Lysis Reagent (Qiagen) combined with the RNeasy Mini Kit (Qiagen).

    Mouse Assay:

    Article Title: Infection- and procedure-dependent effects on pulmonary gene expression in the early phase of influenza A virus infection in mice
    Article Snippet: Untreated mice were used as t = 0 h control. .. Lungs were removed from the cadavers by cutting the main bronchus and were washed in RNAlater RNA Stabilization Reagent (Qiagen Inc, Valencia, CA, USA) immediately after removal.

    Article Title: MicroRNAs Induce Epigenetic Reprogramming and Suppress Malignant Phenotypes of Human Colon Cancer Cells
    Article Snippet: The tumorigenicity assay was performed using an in vivo xenograft model. First, 5 × 106 cells suspended in a total volume of 200 μL DMEM/Matrigel (1:1 (v/v) suspension) were injected into the flanks of 7-8-week-old female mice (BALB/cAJcl-nu/nu). .. Xenograft tumors and mouse blood were collected during sacrifice and preserved in 10% neutral-buffered formalin for histology and immunohistochemical studies or in RNAlater RNA Stabilization Reagent (Qiagen, Tokyo, Japan).

    Article Title: The Absence of DHHC3 Affects Primary and Latent Herpes Simplex Virus 1 Infection
    Article Snippet: Paragraph title: Isolation of RNA from corneas and TG of infected mice. ... Isolated tissues were immersed in RNAlater RNA stabilization reagent (Qiagen, Germantown, MD) and stored at −80°C until they were processed.

    Article Title: Platelet-derived β2M regulates monocyte inflammatory responses
    Article Snippet: .. Hearts from mice were collected day 0 or day 3 after MI and placed into RNAlater RNA Stabilization Reagent (Qiagen). .. Isolated hearts were homogenized using a Tissue-Tearor (BioSpec).

    Activated Clotting Time Assay:

    Article Title: The pea aphid uses a version of the terminal system during oviparous, but not viviparous, development
    Article Snippet: The tissue was placed in RNAlater RNA stabilization reagent (Qiagen, Germantown, USA) and stored at 4°C. .. Real-time qRT-PCR for pea aphid tor was performed on a Step One Plus real-time thermocycler (Life Technologies, Grand Island, USA) using Taqman Gene Expression Master Mix (Life Technologies, Grand Island, USA) and a PrimeTime qPCR Assay (Integrated DNA Technologies, Coralville, USA) that spanned an intron-exon junction (probe 5'-/56-FAM/CAG CAT CAA /ZEN/ATC GTG GTA CGC CAA C/3IABkFQ/-3' with primers 5'-ATT CGA CTC CCT TGC TAT TCG-3' and 5'-TGG GTG ACT TGC AGA CTT AC-3').

    Purification:

    Article Title: The pea aphid uses a version of the terminal system during oviparous, but not viviparous, development
    Article Snippet: The tissue was placed in RNAlater RNA stabilization reagent (Qiagen, Germantown, USA) and stored at 4°C. .. The RNA was purified using the SV Total RNA Isolation System (Promega, Madison, USA) and cDNA was made using the High Capacity cDNA Reverse Transcription Kit (Life Technologies, Grand Island, USA).

    Real-time Polymerase Chain Reaction:

    Article Title: Celastrol inhibits aminoglycoside-induced ototoxicity via heat shock protein 32
    Article Snippet: Control and celastrol-treated utricles were stored in RNAlater RNA Stabilization Reagent (Qiagen, Valencia, CA, USA). .. RNA was reverse transcribed using random hexamer primers (TaqMan, Applied Biosystems), and the resulting cDNA was used for SYBR Green (Applied Biosystems) real-time PCR amplification of HSP70, HSP32, HSP90, HSP27, Nrf2, Gstm1, NQO1, UGT, and GCLC.

    Article Title: Platelet-derived β2M regulates monocyte inflammatory responses
    Article Snippet: Paragraph title: qPCR. ... Hearts from mice were collected day 0 or day 3 after MI and placed into RNAlater RNA Stabilization Reagent (Qiagen).

    Article Title: A Network Analysis of the Human T-Cell Activation Gene Network Identifies Jagged1 as a Therapeutic Target for Autoimmune Diseases
    Article Snippet: RNA extraction, probes and rtPCR Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque (Pharmacia Biotech) and they were immediately submerged in RNAlater RNA Stabilization Reagent (Qiagen) to preserve the gene expression patterns. .. Quantitative real-time PCR (rt-PCR) was performed with the DNA Engine Opticon2 (MJ Research).

    Article Title: The pea aphid uses a version of the terminal system during oviparous, but not viviparous, development
    Article Snippet: The tissue was placed in RNAlater RNA stabilization reagent (Qiagen, Germantown, USA) and stored at 4°C. .. Real-time qRT-PCR for pea aphid tor was performed on a Step One Plus real-time thermocycler (Life Technologies, Grand Island, USA) using Taqman Gene Expression Master Mix (Life Technologies, Grand Island, USA) and a PrimeTime qPCR Assay (Integrated DNA Technologies, Coralville, USA) that spanned an intron-exon junction (probe 5'-/56-FAM/CAG CAT CAA /ZEN/ATC GTG GTA CGC CAA C/3IABkFQ/-3' with primers 5'-ATT CGA CTC CCT TGC TAT TCG-3' and 5'-TGG GTG ACT TGC AGA CTT AC-3').

    Article Title: Immune-Related Gene Expression Profile in Laboratory Common Marmosets Assessed by an Accurate Quantitative Real-Time PCR Using Selected Reference Genes
    Article Snippet: Tissue samples (spleen, mesenteric lymph node, jejunum, ileum, descending colon, cerebrum, cerebellum, brainstem, heart, lung, liver and kidney) were excised from each animal and immediately submerged in RNAlater® RNA Stabilization Reagent (Qiagen). .. All RNA samples were confirmed to have no degradation and were of optimal quality for downstream qPCR applications.

    RNA Extraction:

    Article Title: The Absence of DHHC3 Affects Primary and Latent Herpes Simplex Virus 1 Infection
    Article Snippet: Isolated tissues were immersed in RNAlater RNA stabilization reagent (Qiagen, Germantown, MD) and stored at −80°C until they were processed. .. The corneas or TG from each animal were processed for RNA extraction using TRIzol reagent as we described previously ( ).

    Article Title: A Network Analysis of the Human T-Cell Activation Gene Network Identifies Jagged1 as a Therapeutic Target for Autoimmune Diseases
    Article Snippet: .. RNA extraction, probes and rtPCR Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque (Pharmacia Biotech) and they were immediately submerged in RNAlater RNA Stabilization Reagent (Qiagen) to preserve the gene expression patterns. .. Total RNA was isolated using the RNeasy Mini Kit (Qiagen), removing DNA with the RNase-Free DNase Set (Qiagen), and the High-Capacity cDNA Archive Kit (Applied Biosystems) was used to synthesise cDNA from the total RNA.

    Article Title: Expression Pattern of Peroxisome Proliferator-Activated Receptors in Rat Hippocampus following Cerebral Ischemia and Reperfusion Injury
    Article Snippet: .. Reagents The following reagents were obtained commercially: RNAlater RNA stabilization reagent (Qiagen, Germany); BIOZOL total RNA extraction kit (BioFlux, Japan); ReverTra Ace-α reverse transcription kit (TOYOBO, Japan); mouse anti-rat PPARα , β , and γ monoclonal antibodies (1 : 1000) (Abcam, England); mouse anti-rat superoxide dismutase 2 (SOD2) and mitochondrial uncoupling protein 2 (UCP2) monoclonal antibodies (1 : 1000) (Beijing Biosynthesis Biotechnology, LTD, China); BCA (bicinchoninic acid) protein detection kit (Shanghai Biocolors, China); ECL chemiluminescence detection kit (Pierce Biotech., USA); Taq DNA Polymerase (Promega, USA). ..

    Sample Prep:

    Article Title: Infection- and procedure-dependent effects on pulmonary gene expression in the early phase of influenza A virus infection in mice
    Article Snippet: Paragraph title: Sample preparation ... Lungs were removed from the cadavers by cutting the main bronchus and were washed in RNAlater RNA Stabilization Reagent (Qiagen Inc, Valencia, CA, USA) immediately after removal.

    In Situ:

    Article Title: A Frameshift Mutation within LAMC2 Is Responsible for Herlitz Type Junctional Epidermolysis Bullosa (HJEB) in Black Headed Mutton Sheep
    Article Snippet: .. The samples were harvested immediately after euthanasia and asserved in RNAlater RNA Stabilization Reagent (Qiagen) for stabilization and protection of cellular RNA in situ and then stored at −20°C. .. The RNA was extracted from tissue samples using the Nucleospin RNAII-kit (Macherey-Nagel, Düren, Germany) and transcribed into cDNA using SuperScript III Reverse Transcriptase (Invitrogen, Karlsruhe, Germany).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Changes in Liver Gene Expression and Plasma Concentration of Rbp4, Fetuin-A, and Fgf21 in Sprague-Dawley Rats Subjected to Different Dietary Interventions and Bariatric Surgery
    Article Snippet: Before surgery, reference biopsies of liver tissue were collected, rinsed with PBS, and placed into RNAlater RNA Stabilization Reagent (Qiagen, Hilden, Germany). .. Hepatokines, such as RBP4, fetuin-A, and FGF21, were assessed in duplicate by ELISA kits (Cloud-Clone Corp., Katy, Tex., USA).

    Incubation:

    Article Title: Celastrol inhibits aminoglycoside-induced ototoxicity via heat shock protein 32
    Article Snippet: Real-time quantitative RT-PCR For analysis of celastrol-induced changes in mRNA expression, utricles (three per condition) were incubated in 1.5 μ M. celastrol for 3 h and allowed to recover for 0, 3, 5, 8, or 11 h in culture medium. .. Control and celastrol-treated utricles were stored in RNAlater RNA Stabilization Reagent (Qiagen, Valencia, CA, USA).

    Article Title: Immune-Related Gene Expression Profile in Laboratory Common Marmosets Assessed by an Accurate Quantitative Real-Time PCR Using Selected Reference Genes
    Article Snippet: Following incubation on ice for 5 min, cells were centrifuged at 300×g for 10 min at 4°C and washed with lysis buffer and then PBS. .. Tissue samples (spleen, mesenteric lymph node, jejunum, ileum, descending colon, cerebrum, cerebellum, brainstem, heart, lung, liver and kidney) were excised from each animal and immediately submerged in RNAlater® RNA Stabilization Reagent (Qiagen).

    Sampling:

    Article Title: Changes in Liver Gene Expression and Plasma Concentration of Rbp4, Fetuin-A, and Fgf21 in Sprague-Dawley Rats Subjected to Different Dietary Interventions and Bariatric Surgery
    Article Snippet: After blood sampling the tissues were harvested and the animals were euthanized. .. Liver tissue was explanted, rinsed with PBS, and placed into RNAlater RNA Stabilization Reagent (Qiagen, Hilden, Germany).

    Concentration Assay:

    Article Title: Immune-Related Gene Expression Profile in Laboratory Common Marmosets Assessed by an Accurate Quantitative Real-Time PCR Using Selected Reference Genes
    Article Snippet: Tissue samples (spleen, mesenteric lymph node, jejunum, ileum, descending colon, cerebrum, cerebellum, brainstem, heart, lung, liver and kidney) were excised from each animal and immediately submerged in RNAlater® RNA Stabilization Reagent (Qiagen). .. RNA concentration and integrity were assessed using the Agilent RNA 6,000 Nano Kit (Agilent Technologies, Inc., CA, USA) in an Agilent 2100 Bioanalyzer.

    Marker:

    Article Title: MicroRNAs Induce Epigenetic Reprogramming and Suppress Malignant Phenotypes of Human Colon Cancer Cells
    Article Snippet: Sections were to confirm vasculaturization by immunohistochemistry using CD31, a vascular endothelial marker. .. Xenograft tumors and mouse blood were collected during sacrifice and preserved in 10% neutral-buffered formalin for histology and immunohistochemical studies or in RNAlater RNA Stabilization Reagent (Qiagen, Tokyo, Japan).

    Lysis:

    Article Title: Platelet-derived β2M regulates monocyte inflammatory responses
    Article Snippet: Hearts from mice were collected day 0 or day 3 after MI and placed into RNAlater RNA Stabilization Reagent (Qiagen). .. Isolated primary mouse monocytes from peripheral blood, whole blood platelets, and/or primary heart fibroblasts were pelleted and resuspended in RLT lysis buffer.

    Article Title: Immune-Related Gene Expression Profile in Laboratory Common Marmosets Assessed by an Accurate Quantitative Real-Time PCR Using Selected Reference Genes
    Article Snippet: Leukocytes were lysed with QIAzol® Lysis Reagent (Qiagen, Hilden, Germany) and total RNA was extracted using an RNeasy® Plus Universal Mini Kit (Qiagen) according to the manufacturer's instructions. .. Tissue samples (spleen, mesenteric lymph node, jejunum, ileum, descending colon, cerebrum, cerebellum, brainstem, heart, lung, liver and kidney) were excised from each animal and immediately submerged in RNAlater® RNA Stabilization Reagent (Qiagen).

    Article Title: Concomitant Interferon Alpha Stimulation and TLR3 Activation Induces Neuronal Expression of Depression-Related Genes That Are Elevated in the Brain of Suicidal Persons
    Article Snippet: For RNA protection, the brain tissue was transferred immediately into 1 mL RNAlater RNA Stabilization Reagent (Qiagen) and stored over night at 4°C and then at −20°C. .. Subsequently, total RNA was isolated according to the manufacturer's recommendations for lipid tissues using Qiazol Lysis Reagent (Qiagen) combined with the RNeasy Mini Kit (Qiagen).

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    Qiagen qiagen rnalaterr
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