rnalater  (Qiagen)

 
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    RNAlater RNA Stabilization Reagent
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    For immediate stabilization of RNA in tissues Kit contents Qiagen RNAlater RNA Stabilization Reagent 50mL For Immediate RNA Stabilization and Protection Reliable Gene Expression and Gene profiling Data Protects the RNA Expression Pattern of Samples During Harvest and Storage Preserves RNA for Up to 1 day at 37C 7 days at 18 to 25C or 4 Weeks at 2 to 8C Placed at 20C or 80C for Archival Storage Ideal for Total RNA miRNA mRNA Purification Benefits Immediate RNA stabilization and protection Reliable gene expression and gene profiling data Convenient and safe handling at room temperature No need for liquid nitrogen or dry ice Tissue archiving without risk of RNA degrad
    Catalog Number:
    76104
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    RNAlater RNA Stabilization Reagent
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    Qiagen rnalater
    RNAlater RNA Stabilization Reagent
    For immediate stabilization of RNA in tissues Kit contents Qiagen RNAlater RNA Stabilization Reagent 50mL For Immediate RNA Stabilization and Protection Reliable Gene Expression and Gene profiling Data Protects the RNA Expression Pattern of Samples During Harvest and Storage Preserves RNA for Up to 1 day at 37C 7 days at 18 to 25C or 4 Weeks at 2 to 8C Placed at 20C or 80C for Archival Storage Ideal for Total RNA miRNA mRNA Purification Benefits Immediate RNA stabilization and protection Reliable gene expression and gene profiling data Convenient and safe handling at room temperature No need for liquid nitrogen or dry ice Tissue archiving without risk of RNA degrad
    https://www.bioz.com/result/rnalater/product/Qiagen
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    Images

    1) 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

    2) 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

    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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    Isolation:

    Article Title: A subset of chemosensory genes differs between two populations of a specialized leaf beetle after host plant shift, et al. A subset of chemosensory genes differs between two populations of a specialized leaf beetle after host plant shift
    Article Snippet: .. After these 2 days, individuals from C. lapponica were dissected and the organs were stored in RNA stabilization solution (RNAlater, Qiagen, Hilden, Germany) for transport to Germany where RNA isolation took place in the laboratory. ..

    Preserving:

    Article Title: Age-Dependent Decline in Mouse Lung Regeneration with Loss of Lung Fibroblast Clonogenicity and Increased Myofibroblastic Differentiation
    Article Snippet: .. RNA preservation was achieved by flooding the lung intratracheally with RNAlater solution (Qiagen #76104), followed by storage of lung tissue samples in RNAlater at −80°C. .. The control samples used in this microarray experiment were the left lungs obtained from the study animals at the time of PNX and stored in RNAlater at −80°C.

    RNA Sequencing Assay:

    Article Title: Transcriptome Characterization by RNA-Seq Reveals the Involvement of the Complement Components in Noise-Traumatized Rat Cochleae
    Article Snippet: .. The cochleae to be used for RNA-seq and qRT-PCR analysis were immediately perfused through the round window with an RNA stabilization reagent (RNAlater, Qiagen, Valencia, CA). .. The cochlea was then carefully dissected to collect cochlear tissues in the RNAlater reagent under a dissection microscope.

    Expressing:

    Article Title: Enhancing tristetraprolin activity reduces the severity of cigarette smoke‐induced experimental chronic obstructive pulmonary disease
    Article Snippet: .. mRNA expression Whole lungs were collected and stored in RNA Stabilization Reagent, RNAlater (Qiagen, Chadstone Centre, Vic, Australia). ..

    Quantitative RT-PCR:

    Article Title: Transcriptome Characterization by RNA-Seq Reveals the Involvement of the Complement Components in Noise-Traumatized Rat Cochleae
    Article Snippet: .. The cochleae to be used for RNA-seq and qRT-PCR analysis were immediately perfused through the round window with an RNA stabilization reagent (RNAlater, Qiagen, Valencia, CA). .. The cochlea was then carefully dissected to collect cochlear tissues in the RNAlater reagent under a dissection microscope.

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