enscs  (Qiagen)


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    Name:
    RNeasy Plus Micro Kit
    Description:
    For purification of up to 45 µg total RNA from cells tissues using gDNA Eliminator columns Kit contents Qiagen RNeasy Plus Micro Kit 50 preps 14L Elution Volume 5mg Tissue Amount Tissue Cells Sample Total RNA Purification Spin Column Format Silica Technology Ideal for Northern Dot and Slot Blotting End point RT PCR Quantitative Real time RT PCR Includes RNeasy MinElute Spin Columns gDNA Eliminator Spin Columns Collection Tubes Carrier RNA RNase free Water and Buffers Benefits Unique gDNA Eliminator columns avoid the need for DNase Efficient removal of genomic DNA Highly reproducible yields of RNA in minutes High performance RNA for sensitive application
    Catalog Number:
    74034
    Price:
    436
    Category:
    RNeasy Plus Micro Kit
    Buy from Supplier


    Structured Review

    Qiagen enscs
    RNeasy Plus Micro Kit
    For purification of up to 45 µg total RNA from cells tissues using gDNA Eliminator columns Kit contents Qiagen RNeasy Plus Micro Kit 50 preps 14L Elution Volume 5mg Tissue Amount Tissue Cells Sample Total RNA Purification Spin Column Format Silica Technology Ideal for Northern Dot and Slot Blotting End point RT PCR Quantitative Real time RT PCR Includes RNeasy MinElute Spin Columns gDNA Eliminator Spin Columns Collection Tubes Carrier RNA RNase free Water and Buffers Benefits Unique gDNA Eliminator columns avoid the need for DNase Efficient removal of genomic DNA Highly reproducible yields of RNA in minutes High performance RNA for sensitive application
    https://www.bioz.com/result/enscs/product/Qiagen
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    enscs - by Bioz Stars, 2021-03
    97/100 stars

    Images

    1) Product Images from "Effect of Bmi1 over-expression on gene expression in adult and embryonic murine neural stem cells"

    Article Title: Effect of Bmi1 over-expression on gene expression in adult and embryonic murine neural stem cells

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-25921-8

    Effect on gene expression of Bmi1 overexpression in eNSCs compared to aNSCs. ( A ) Comparison of transcript levels measured by RNA-seq (reads per kilobase per million reads mapped, RPKM) in embryonic and adult NSCs (empty vector control, average of two biological replicates). ( B ) Gene ontology categories enriched for genes down-regulated (top) or up-regulated (bottom) upon Bmi1 overexpression in eNSCs (red bars) or aNSCs (blue bars). Categories having P
    Figure Legend Snippet: Effect on gene expression of Bmi1 overexpression in eNSCs compared to aNSCs. ( A ) Comparison of transcript levels measured by RNA-seq (reads per kilobase per million reads mapped, RPKM) in embryonic and adult NSCs (empty vector control, average of two biological replicates). ( B ) Gene ontology categories enriched for genes down-regulated (top) or up-regulated (bottom) upon Bmi1 overexpression in eNSCs (red bars) or aNSCs (blue bars). Categories having P

    Techniques Used: Expressing, Over Expression, RNA Sequencing Assay, Plasmid Preparation

    Comparison of gene expression changes in embryonic and adult NSCs caused by Bmi1 overexpression. ( A ) Reads per kilobase per million reads (RPKM) from RNA-seq data from aNSCs and eSNCs is plotted for 4296 genes having expression altered at least two-fold by Bmi1 overexpression in either eNSCs or aNSCs. ( B ) Genes plotted in ( A ) were broken down according to their relative expression in eNSCs and aNSCs as indicated.
    Figure Legend Snippet: Comparison of gene expression changes in embryonic and adult NSCs caused by Bmi1 overexpression. ( A ) Reads per kilobase per million reads (RPKM) from RNA-seq data from aNSCs and eSNCs is plotted for 4296 genes having expression altered at least two-fold by Bmi1 overexpression in either eNSCs or aNSCs. ( B ) Genes plotted in ( A ) were broken down according to their relative expression in eNSCs and aNSCs as indicated.

    Techniques Used: Expressing, Over Expression, RNA Sequencing Assay

    2) Product Images from "Filaggrin deficient mice have a lower threshold for cutaneous allergen sensitization but do not develop spontaneous skin inflammation or atopy"

    Article Title: Filaggrin deficient mice have a lower threshold for cutaneous allergen sensitization but do not develop spontaneous skin inflammation or atopy

    Journal: bioRxiv

    doi: 10.1101/2020.09.11.293688

    Alteration of keratinocyte gene expression and composition of microbiota of Flg -/- skin. A) RNA sequencing analysis of basal CD49f + keratinocytes (negative for CD45, see also Fig S3), sorted from skin cell suspensions of four 8-12 week-old female Flg -/- mice and 5 littermate controls. Heat map represents expression of genes involved in antigen presentation and the Retnla gene (relative expression, individual read counts corrected for the mean expression of the respective gene), p
    Figure Legend Snippet: Alteration of keratinocyte gene expression and composition of microbiota of Flg -/- skin. A) RNA sequencing analysis of basal CD49f + keratinocytes (negative for CD45, see also Fig S3), sorted from skin cell suspensions of four 8-12 week-old female Flg -/- mice and 5 littermate controls. Heat map represents expression of genes involved in antigen presentation and the Retnla gene (relative expression, individual read counts corrected for the mean expression of the respective gene), p

    Techniques Used: Expressing, RNA Sequencing Assay, Mouse Assay

    3) Product Images from "Cholinergic signals from the CNS regulate G-CSF-mediated HSC mobilization from bone marrow via a glucocorticoid signaling relay"

    Article Title: Cholinergic signals from the CNS regulate G-CSF-mediated HSC mobilization from bone marrow via a glucocorticoid signaling relay

    Journal: Cell stem cell

    doi: 10.1016/j.stem.2017.01.002

    G-CSF-induced HSC mobilization requires signals from the muscarinic receptor type-1 ( Chrm1 ) (A) HSCs/mL following G-CSF mobilization in wild-type mice treated with saline or Scopolamine hydrobromide (Scop 1mg/kg; n = 5–8). HSCs defined as Lineage − Sca1 + cKit + Flt3 − . (B) Schematic of G-CSF-induced mobilization and analyses. Wild-type mice are denoted by black bars and Chrm1 −/− mice by white bars. (C) HSCs/ml of peripheral blood (Lineage − Sca1 + cKit + Flt3 − ; n = 12–16) ( D) Colony-forming units/ml of peripheral blood determined in vitro (CFU-C; n = 6–13). (E) Lineage − Sca1 + cKit + (LSK) cells/femur at steady state (n = 9). ( F) Reconstitution after competitive BM transplantation of mobilized blood into irradiated hosts ( G) Tri-lineage engraftment of donor cells 16 weeks post transplantation: B220 + cells (B) CD4/CD8 + cells (T) and Mac1 + cells (M; n= 6–7). * p
    Figure Legend Snippet: G-CSF-induced HSC mobilization requires signals from the muscarinic receptor type-1 ( Chrm1 ) (A) HSCs/mL following G-CSF mobilization in wild-type mice treated with saline or Scopolamine hydrobromide (Scop 1mg/kg; n = 5–8). HSCs defined as Lineage − Sca1 + cKit + Flt3 − . (B) Schematic of G-CSF-induced mobilization and analyses. Wild-type mice are denoted by black bars and Chrm1 −/− mice by white bars. (C) HSCs/ml of peripheral blood (Lineage − Sca1 + cKit + Flt3 − ; n = 12–16) ( D) Colony-forming units/ml of peripheral blood determined in vitro (CFU-C; n = 6–13). (E) Lineage − Sca1 + cKit + (LSK) cells/femur at steady state (n = 9). ( F) Reconstitution after competitive BM transplantation of mobilized blood into irradiated hosts ( G) Tri-lineage engraftment of donor cells 16 weeks post transplantation: B220 + cells (B) CD4/CD8 + cells (T) and Mac1 + cells (M; n= 6–7). * p

    Techniques Used: Mouse Assay, In Vitro, Transplantation Assay, Irradiation

    4) Product Images from "Effect of Bmi1 over-expression on gene expression in adult and embryonic murine neural stem cells"

    Article Title: Effect of Bmi1 over-expression on gene expression in adult and embryonic murine neural stem cells

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-25921-8

    Effect on gene expression of Bmi1 overexpression in eNSCs compared to aNSCs. ( A ) Comparison of transcript levels measured by RNA-seq (reads per kilobase per million reads mapped, RPKM) in embryonic and adult NSCs (empty vector control, average of two biological replicates). ( B ) Gene ontology categories enriched for genes down-regulated (top) or up-regulated (bottom) upon Bmi1 overexpression in eNSCs (red bars) or aNSCs (blue bars). Categories having P
    Figure Legend Snippet: Effect on gene expression of Bmi1 overexpression in eNSCs compared to aNSCs. ( A ) Comparison of transcript levels measured by RNA-seq (reads per kilobase per million reads mapped, RPKM) in embryonic and adult NSCs (empty vector control, average of two biological replicates). ( B ) Gene ontology categories enriched for genes down-regulated (top) or up-regulated (bottom) upon Bmi1 overexpression in eNSCs (red bars) or aNSCs (blue bars). Categories having P

    Techniques Used: Expressing, Over Expression, RNA Sequencing Assay, Plasmid Preparation

    Comparison of gene expression changes in embryonic and adult NSCs caused by Bmi1 overexpression. ( A ) Reads per kilobase per million reads (RPKM) from RNA-seq data from aNSCs and eSNCs is plotted for 4296 genes having expression altered at least two-fold by Bmi1 overexpression in either eNSCs or aNSCs. ( B ) Genes plotted in ( A ) were broken down according to their relative expression in eNSCs and aNSCs as indicated.
    Figure Legend Snippet: Comparison of gene expression changes in embryonic and adult NSCs caused by Bmi1 overexpression. ( A ) Reads per kilobase per million reads (RPKM) from RNA-seq data from aNSCs and eSNCs is plotted for 4296 genes having expression altered at least two-fold by Bmi1 overexpression in either eNSCs or aNSCs. ( B ) Genes plotted in ( A ) were broken down according to their relative expression in eNSCs and aNSCs as indicated.

    Techniques Used: Expressing, Over Expression, RNA Sequencing Assay

    Related Articles

    Quantitative RT-PCR:

    Article Title: Anti-fibrotic action of pirfenidone in Dupuytren’s disease-derived fibroblasts
    Article Snippet: .. Quantitative Real-time RT-PCR (qRT-PCR) Total RNA isolated (RNeasy Micro Kit, Qiagen Inc, Valencia, CA) from CT- and DD-derived fibroblasts treated and untreated with PFD (800 μg/ml) and TGF-β1 (10 ng/ml) was subjected to real-time RT-PCR to determine the mRNA expression levels for α-SMA (ACTA2 ), type I collagen (COL1A2 ), and type III collagen (COL3A1 ). ..

    Article Title: Filaggrin deficient mice have a lower threshold for cutaneous allergen sensitization but do not develop spontaneous skin inflammation or atopy
    Article Snippet: RNAseq differential expression analysis was performed with the R package DESeq2 ( ). .. quantitative RT-PCR Total RNA from sorted CD45-negative cells was isolated using RNeasy® Plus Micro Kit (Qiagen), while RNA from whole ear skin tissue was extracted using RNeasy® Mini Kit (Qiagen), according to the manufacturer’s instructions. cDNA was synthesized using the PrimeScript RT Reagent Kit (Takara). .. Quantification of transcripts was performed on a Mx3005P QPCR system (Agilent Technologies) for the IL-1b expression assay, using Maxima SYBR green/ROX qPCR Master Mix (Thermo Fisher Scientific).

    Isolation:

    Article Title: Anti-fibrotic action of pirfenidone in Dupuytren’s disease-derived fibroblasts
    Article Snippet: .. Quantitative Real-time RT-PCR (qRT-PCR) Total RNA isolated (RNeasy Micro Kit, Qiagen Inc, Valencia, CA) from CT- and DD-derived fibroblasts treated and untreated with PFD (800 μg/ml) and TGF-β1 (10 ng/ml) was subjected to real-time RT-PCR to determine the mRNA expression levels for α-SMA (ACTA2 ), type I collagen (COL1A2 ), and type III collagen (COL3A1 ). ..

    Article Title: Filaggrin deficient mice have a lower threshold for cutaneous allergen sensitization but do not develop spontaneous skin inflammation or atopy
    Article Snippet: RNAseq differential expression analysis was performed with the R package DESeq2 ( ). .. quantitative RT-PCR Total RNA from sorted CD45-negative cells was isolated using RNeasy® Plus Micro Kit (Qiagen), while RNA from whole ear skin tissue was extracted using RNeasy® Mini Kit (Qiagen), according to the manufacturer’s instructions. cDNA was synthesized using the PrimeScript RT Reagent Kit (Takara). .. Quantification of transcripts was performed on a Mx3005P QPCR system (Agilent Technologies) for the IL-1b expression assay, using Maxima SYBR green/ROX qPCR Master Mix (Thermo Fisher Scientific).

    Article Title: Spinal neurons require Islet1 for subtype-specific differentiation of electrical excitability
    Article Snippet: Control or E3MO sorted cell samples in Lysis Buffer were pooled in groups corresponding to a total of 80,000 to 100,000 cells for total RNA extraction (control groups, n = 5; E3MO groups, n = 3). .. Real time quantitative PCR Total RNA was extracted from FAC sorted GFP+ cells using RNA column-based isolation kits, RNeasy® Micro kit (Qiagen). .. Concentration and integrity of the RNA extracted from FAC sorted cells were determined with an Agilent 2100 Bioanalyzer and by use of the Agilent RNA 6000 Pico Kit (Agilent Technologies, Santa Clara, CA, USA).

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes
    Article Snippet: The RNAqueous micro kit and RNeasy plus micro kit enable high quality RNA purification from low numbers of FACS sorted EGFP positive zebrafish cells In search for an appropriate method to isolate RNA from a low number (5000–200,000) of FACS sorted zebrafish cells, two RNA isolation kits were compared. .. The RNAqueous Micro Total RNA Isolation Kit from Ambion and the RNeasy Plus Micro Kit from Qiagen are specifically designed for purification of total RNA from a small amount of cells ( < 200,000 cells). .. In both cases, only longer RNA fragments ( > 200 nucleotides) are purified, although the workflow can be modified to isolate small RNAs such as microRNAs, 5,8S rRNA, 5S RNA, tRNA’s,… (Additional file : Figure S1).

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes
    Article Snippet: We compared the performance of two RNA isolation kits for obtaining high-quality RNA from FACS sorted cells, i.e. the RNAqueous Micro Total RNA Isolation Kit from Ambion and the RNeasy Plus Micro Kit from Qiagen. .. Based on different quality parameters and comparison of various experimental set-ups, we recommend using the RNeasy Plus Micro Kit for RNA isolation of FACS sorted zebrafish cells. .. The RNA purification protocol is fast (only 20–30 min), easy and consistently delivers high-quality RNA samples.

    Article Title: Nicotinamide Ameliorates Disease Phenotypes in a Human iPSC Model of Age-related Macular Degeneration
    Article Snippet: G-band karyotyping was performed by the Cytogenetics laboratory of the Wadsworth Center at the David Axelrod Institute (Albany, NY), and by Cell Guidance Systems Genetics Service (St. Louis, MO). .. To confirm that no residual RPE donor cells contaminated the hiPSC cultures, total RNA was isolated (Cat. # 74034; Qiagen) from each hiPSC line, cDNA was reverse transcribed (Cat. # 4387406; Thermo Fisher) and qPCR performed using primers specific for the RPE gene RPE65 ( ). .. hiPSC lines were grown on irradiated mouse embryonic feeders (Cat. # GSE-6301G; GlobalStem) in serum-free medium supplemented with 4ng/ml FGF2 (Cat. # 233-FB; R & D systems).

    Expressing:

    Article Title: Anti-fibrotic action of pirfenidone in Dupuytren’s disease-derived fibroblasts
    Article Snippet: .. Quantitative Real-time RT-PCR (qRT-PCR) Total RNA isolated (RNeasy Micro Kit, Qiagen Inc, Valencia, CA) from CT- and DD-derived fibroblasts treated and untreated with PFD (800 μg/ml) and TGF-β1 (10 ng/ml) was subjected to real-time RT-PCR to determine the mRNA expression levels for α-SMA (ACTA2 ), type I collagen (COL1A2 ), and type III collagen (COL3A1 ). ..

    Synthesized:

    Article Title: Filaggrin deficient mice have a lower threshold for cutaneous allergen sensitization but do not develop spontaneous skin inflammation or atopy
    Article Snippet: RNAseq differential expression analysis was performed with the R package DESeq2 ( ). .. quantitative RT-PCR Total RNA from sorted CD45-negative cells was isolated using RNeasy® Plus Micro Kit (Qiagen), while RNA from whole ear skin tissue was extracted using RNeasy® Mini Kit (Qiagen), according to the manufacturer’s instructions. cDNA was synthesized using the PrimeScript RT Reagent Kit (Takara). .. Quantification of transcripts was performed on a Mx3005P QPCR system (Agilent Technologies) for the IL-1b expression assay, using Maxima SYBR green/ROX qPCR Master Mix (Thermo Fisher Scientific).

    Real-time Polymerase Chain Reaction:

    Article Title: Spinal neurons require Islet1 for subtype-specific differentiation of electrical excitability
    Article Snippet: Control or E3MO sorted cell samples in Lysis Buffer were pooled in groups corresponding to a total of 80,000 to 100,000 cells for total RNA extraction (control groups, n = 5; E3MO groups, n = 3). .. Real time quantitative PCR Total RNA was extracted from FAC sorted GFP+ cells using RNA column-based isolation kits, RNeasy® Micro kit (Qiagen). .. Concentration and integrity of the RNA extracted from FAC sorted cells were determined with an Agilent 2100 Bioanalyzer and by use of the Agilent RNA 6000 Pico Kit (Agilent Technologies, Santa Clara, CA, USA).

    Article Title: Nicotinamide Ameliorates Disease Phenotypes in a Human iPSC Model of Age-related Macular Degeneration
    Article Snippet: G-band karyotyping was performed by the Cytogenetics laboratory of the Wadsworth Center at the David Axelrod Institute (Albany, NY), and by Cell Guidance Systems Genetics Service (St. Louis, MO). .. To confirm that no residual RPE donor cells contaminated the hiPSC cultures, total RNA was isolated (Cat. # 74034; Qiagen) from each hiPSC line, cDNA was reverse transcribed (Cat. # 4387406; Thermo Fisher) and qPCR performed using primers specific for the RPE gene RPE65 ( ). .. hiPSC lines were grown on irradiated mouse embryonic feeders (Cat. # GSE-6301G; GlobalStem) in serum-free medium supplemented with 4ng/ml FGF2 (Cat. # 233-FB; R & D systems).

    Purification:

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes
    Article Snippet: The RNAqueous micro kit and RNeasy plus micro kit enable high quality RNA purification from low numbers of FACS sorted EGFP positive zebrafish cells In search for an appropriate method to isolate RNA from a low number (5000–200,000) of FACS sorted zebrafish cells, two RNA isolation kits were compared. .. The RNAqueous Micro Total RNA Isolation Kit from Ambion and the RNeasy Plus Micro Kit from Qiagen are specifically designed for purification of total RNA from a small amount of cells ( < 200,000 cells). .. In both cases, only longer RNA fragments ( > 200 nucleotides) are purified, although the workflow can be modified to isolate small RNAs such as microRNAs, 5,8S rRNA, 5S RNA, tRNA’s,… (Additional file : Figure S1).

    FACS:

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes
    Article Snippet: We compared the performance of two RNA isolation kits for obtaining high-quality RNA from FACS sorted cells, i.e. the RNAqueous Micro Total RNA Isolation Kit from Ambion and the RNeasy Plus Micro Kit from Qiagen. .. Based on different quality parameters and comparison of various experimental set-ups, we recommend using the RNeasy Plus Micro Kit for RNA isolation of FACS sorted zebrafish cells. .. The RNA purification protocol is fast (only 20–30 min), easy and consistently delivers high-quality RNA samples.

    High Throughput Screening Assay:

    Article Title: Effect of Bmi1 over-expression on gene expression in adult and embryonic murine neural stem cells
    Article Snippet: .. High throughput RNA sequencing (RNA-seq) To prepare samples for Illumina sequencing, 150–250 ng of RNA prepared from aNSCs or eNSCs (using the RNAeasy Plus Micro kit; Qiagen) was first treated to remove ribosomal RNA using the Ribo-Zero Magnetic Kit (Epicentre, Madison, WI), using the protocol for low input samples. .. Sequencing libraries were prepared using the ScriptSeq V2 RNA-seq Kit (Epicentre), again using the low input protocol.

    RNA Sequencing Assay:

    Article Title: Effect of Bmi1 over-expression on gene expression in adult and embryonic murine neural stem cells
    Article Snippet: .. High throughput RNA sequencing (RNA-seq) To prepare samples for Illumina sequencing, 150–250 ng of RNA prepared from aNSCs or eNSCs (using the RNAeasy Plus Micro kit; Qiagen) was first treated to remove ribosomal RNA using the Ribo-Zero Magnetic Kit (Epicentre, Madison, WI), using the protocol for low input samples. .. Sequencing libraries were prepared using the ScriptSeq V2 RNA-seq Kit (Epicentre), again using the low input protocol.

    Sequencing:

    Article Title: Effect of Bmi1 over-expression on gene expression in adult and embryonic murine neural stem cells
    Article Snippet: .. High throughput RNA sequencing (RNA-seq) To prepare samples for Illumina sequencing, 150–250 ng of RNA prepared from aNSCs or eNSCs (using the RNAeasy Plus Micro kit; Qiagen) was first treated to remove ribosomal RNA using the Ribo-Zero Magnetic Kit (Epicentre, Madison, WI), using the protocol for low input samples. .. Sequencing libraries were prepared using the ScriptSeq V2 RNA-seq Kit (Epicentre), again using the low input protocol.

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    Qiagen rna column based isolation kits
    Islet1 knock-down affects sensory neuron differentiation. (A-C) In control embryos (A, B) , Rohon-Beard cells (RBs) express runx3 . Islet1 knock-down leads to fewer cells with robust expression of runx3 (C). The asterisk denotes a cell expressing the gene. (D-F’) <t>Tg(-3.4neurog1:gfp)sb4</t> control (D-E’) and E3 morphant (F, F’) 24 hours post-fertilization (hpf) embryos were examined for expression of olig4 (red). (D-E’) In lateral views of the dorsal spinal cord, <t>RNA</t> in situ hybridization reveals expression of the interneuron marker olig4 (red). Green fluorescent protein (GFP) + neurons do not express olig4 and comprise RBs and dorsal lateral ascending interneurons (see Figure 2 ). (F-F’) Islet1 knock-down leads to an increase in the number of olig4 expressing cells within the dorsal spinal cord. However, similar to RBs of control embryos (D-E’) , RB-like neurons do not express detectable levels of olig4 (F) . (G-I) At 72 hpf, dorsal root ganglia (DRGs) are easily identified as GFP + neurons with large somata located near the spinal cord/notochord border. (G, H) In control embryos, DRG neurons project from their soma bipolar axons that extend dorsally and ventrally (asterisks). (I) Islet1 knock-down reduces the number of GFP + DRGs. Furthermore, for the few GFP + DRGs remaining, their axons show abnormal morphologies. Scale bars = 50 μm in A (for A-C ), D (for D-F ’) and G (for G-I ). CtlMO, 5-base mismatched; islet1 (Sp)E3MO; E3MO, E3 morpholino; Uninj, uninjected.
    Rna Column Based Isolation Kits, supplied by Qiagen, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna column based isolation kits/product/Qiagen
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rna column based isolation kits - by Bioz Stars, 2021-03
    97/100 stars
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    Islet1 knock-down affects sensory neuron differentiation. (A-C) In control embryos (A, B) , Rohon-Beard cells (RBs) express runx3 . Islet1 knock-down leads to fewer cells with robust expression of runx3 (C). The asterisk denotes a cell expressing the gene. (D-F’) Tg(-3.4neurog1:gfp)sb4 control (D-E’) and E3 morphant (F, F’) 24 hours post-fertilization (hpf) embryos were examined for expression of olig4 (red). (D-E’) In lateral views of the dorsal spinal cord, RNA in situ hybridization reveals expression of the interneuron marker olig4 (red). Green fluorescent protein (GFP) + neurons do not express olig4 and comprise RBs and dorsal lateral ascending interneurons (see Figure 2 ). (F-F’) Islet1 knock-down leads to an increase in the number of olig4 expressing cells within the dorsal spinal cord. However, similar to RBs of control embryos (D-E’) , RB-like neurons do not express detectable levels of olig4 (F) . (G-I) At 72 hpf, dorsal root ganglia (DRGs) are easily identified as GFP + neurons with large somata located near the spinal cord/notochord border. (G, H) In control embryos, DRG neurons project from their soma bipolar axons that extend dorsally and ventrally (asterisks). (I) Islet1 knock-down reduces the number of GFP + DRGs. Furthermore, for the few GFP + DRGs remaining, their axons show abnormal morphologies. Scale bars = 50 μm in A (for A-C ), D (for D-F ’) and G (for G-I ). CtlMO, 5-base mismatched; islet1 (Sp)E3MO; E3MO, E3 morpholino; Uninj, uninjected.

    Journal: Neural Development

    Article Title: Spinal neurons require Islet1 for subtype-specific differentiation of electrical excitability

    doi: 10.1186/1749-8104-9-19

    Figure Lengend Snippet: Islet1 knock-down affects sensory neuron differentiation. (A-C) In control embryos (A, B) , Rohon-Beard cells (RBs) express runx3 . Islet1 knock-down leads to fewer cells with robust expression of runx3 (C). The asterisk denotes a cell expressing the gene. (D-F’) Tg(-3.4neurog1:gfp)sb4 control (D-E’) and E3 morphant (F, F’) 24 hours post-fertilization (hpf) embryos were examined for expression of olig4 (red). (D-E’) In lateral views of the dorsal spinal cord, RNA in situ hybridization reveals expression of the interneuron marker olig4 (red). Green fluorescent protein (GFP) + neurons do not express olig4 and comprise RBs and dorsal lateral ascending interneurons (see Figure 2 ). (F-F’) Islet1 knock-down leads to an increase in the number of olig4 expressing cells within the dorsal spinal cord. However, similar to RBs of control embryos (D-E’) , RB-like neurons do not express detectable levels of olig4 (F) . (G-I) At 72 hpf, dorsal root ganglia (DRGs) are easily identified as GFP + neurons with large somata located near the spinal cord/notochord border. (G, H) In control embryos, DRG neurons project from their soma bipolar axons that extend dorsally and ventrally (asterisks). (I) Islet1 knock-down reduces the number of GFP + DRGs. Furthermore, for the few GFP + DRGs remaining, their axons show abnormal morphologies. Scale bars = 50 μm in A (for A-C ), D (for D-F ’) and G (for G-I ). CtlMO, 5-base mismatched; islet1 (Sp)E3MO; E3MO, E3 morpholino; Uninj, uninjected.

    Article Snippet: Real time quantitative PCR Total RNA was extracted from FAC sorted GFP+ cells using RNA column-based isolation kits, RNeasy® Micro kit (Qiagen).

    Techniques: Expressing, RNA In Situ Hybridization, Marker

    Effect on gene expression of Bmi1 overexpression in eNSCs compared to aNSCs. ( A ) Comparison of transcript levels measured by RNA-seq (reads per kilobase per million reads mapped, RPKM) in embryonic and adult NSCs (empty vector control, average of two biological replicates). ( B ) Gene ontology categories enriched for genes down-regulated (top) or up-regulated (bottom) upon Bmi1 overexpression in eNSCs (red bars) or aNSCs (blue bars). Categories having P

    Journal: Scientific Reports

    Article Title: Effect of Bmi1 over-expression on gene expression in adult and embryonic murine neural stem cells

    doi: 10.1038/s41598-018-25921-8

    Figure Lengend Snippet: Effect on gene expression of Bmi1 overexpression in eNSCs compared to aNSCs. ( A ) Comparison of transcript levels measured by RNA-seq (reads per kilobase per million reads mapped, RPKM) in embryonic and adult NSCs (empty vector control, average of two biological replicates). ( B ) Gene ontology categories enriched for genes down-regulated (top) or up-regulated (bottom) upon Bmi1 overexpression in eNSCs (red bars) or aNSCs (blue bars). Categories having P

    Article Snippet: High throughput RNA sequencing (RNA-seq) To prepare samples for Illumina sequencing, 150–250 ng of RNA prepared from aNSCs or eNSCs (using the RNAeasy Plus Micro kit; Qiagen) was first treated to remove ribosomal RNA using the Ribo-Zero Magnetic Kit (Epicentre, Madison, WI), using the protocol for low input samples.

    Techniques: Expressing, Over Expression, RNA Sequencing Assay, Plasmid Preparation

    Comparison of gene expression changes in embryonic and adult NSCs caused by Bmi1 overexpression. ( A ) Reads per kilobase per million reads (RPKM) from RNA-seq data from aNSCs and eSNCs is plotted for 4296 genes having expression altered at least two-fold by Bmi1 overexpression in either eNSCs or aNSCs. ( B ) Genes plotted in ( A ) were broken down according to their relative expression in eNSCs and aNSCs as indicated.

    Journal: Scientific Reports

    Article Title: Effect of Bmi1 over-expression on gene expression in adult and embryonic murine neural stem cells

    doi: 10.1038/s41598-018-25921-8

    Figure Lengend Snippet: Comparison of gene expression changes in embryonic and adult NSCs caused by Bmi1 overexpression. ( A ) Reads per kilobase per million reads (RPKM) from RNA-seq data from aNSCs and eSNCs is plotted for 4296 genes having expression altered at least two-fold by Bmi1 overexpression in either eNSCs or aNSCs. ( B ) Genes plotted in ( A ) were broken down according to their relative expression in eNSCs and aNSCs as indicated.

    Article Snippet: High throughput RNA sequencing (RNA-seq) To prepare samples for Illumina sequencing, 150–250 ng of RNA prepared from aNSCs or eNSCs (using the RNAeasy Plus Micro kit; Qiagen) was first treated to remove ribosomal RNA using the Ribo-Zero Magnetic Kit (Epicentre, Madison, WI), using the protocol for low input samples.

    Techniques: Expressing, Over Expression, RNA Sequencing Assay

    Sorting small cell populations directly into the lysis buffer of the RNA isolation kit improves RNA quality and yield. ( a ) RQN values ( b ) and RNA yield of RNA samples isolated from a range of cell numbers (5000–200,000) when sorting directly into the lysis buffer of the RNA isolation kit or collecting the cells first into a collection medium. RQN values ( c ) and RNA yield ( d ) of RNA samples isolated from a range of cell numbers (5000–200,000) sorted directly into the lysis buffer or sorted into a collection buffer first. But here the volume of the lysis buffer was amended not to exceed its maximum dilution point caused by the sorting procedure. Left panels show samples isolated with the RNeasy plus micro kit, right panels with the RNAqueous micro kit. Average of two biological replicates is shown. Error bars indicate SEM

    Journal: BMC Genomics

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes

    doi: 10.1186/s12864-019-5608-2

    Figure Lengend Snippet: Sorting small cell populations directly into the lysis buffer of the RNA isolation kit improves RNA quality and yield. ( a ) RQN values ( b ) and RNA yield of RNA samples isolated from a range of cell numbers (5000–200,000) when sorting directly into the lysis buffer of the RNA isolation kit or collecting the cells first into a collection medium. RQN values ( c ) and RNA yield ( d ) of RNA samples isolated from a range of cell numbers (5000–200,000) sorted directly into the lysis buffer or sorted into a collection buffer first. But here the volume of the lysis buffer was amended not to exceed its maximum dilution point caused by the sorting procedure. Left panels show samples isolated with the RNeasy plus micro kit, right panels with the RNAqueous micro kit. Average of two biological replicates is shown. Error bars indicate SEM

    Article Snippet: Based on different quality parameters and comparison of various experimental set-ups, we recommend using the RNeasy Plus Micro Kit for RNA isolation of FACS sorted zebrafish cells.

    Techniques: Lysis, Isolation

    Sorting cells directly into the lysis buffer preserves and protects RNA from degradation. RQN values of individual RNA samples isolated at specific time points after FACS sorting of 20,000 fli1a:EGFP cells. Cells were sorted directly in the lysis buffer of the kit or first captured into a collection medium. Left panel shows samples isolated with the RNAqueous micro kit, right panel with the RNeasy plus micro kit

    Journal: BMC Genomics

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes

    doi: 10.1186/s12864-019-5608-2

    Figure Lengend Snippet: Sorting cells directly into the lysis buffer preserves and protects RNA from degradation. RQN values of individual RNA samples isolated at specific time points after FACS sorting of 20,000 fli1a:EGFP cells. Cells were sorted directly in the lysis buffer of the kit or first captured into a collection medium. Left panel shows samples isolated with the RNAqueous micro kit, right panel with the RNeasy plus micro kit

    Article Snippet: Based on different quality parameters and comparison of various experimental set-ups, we recommend using the RNeasy Plus Micro Kit for RNA isolation of FACS sorted zebrafish cells.

    Techniques: Lysis, Isolation, FACS

    Comparative qualitative analysis of RNA purified with the Rnaqeuous and Rneasy micro kit. ( a ) Boxplots showing RQN values and ( b ) RNA yield of 10 RNA samples purified from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent RQN values of individual samples. ( c ) Boxplots showing 5′-3′ delta-Cq (dCq) values calculated from a 5′ and a 3′ RT-qPCR assay of 7 RNA samples isolated from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent 5′-3′ dCq values of individual samples. * P

    Journal: BMC Genomics

    Article Title: Purification of high-quality RNA from a small number of fluorescence activated cell sorted zebrafish cells for RNA sequencing purposes

    doi: 10.1186/s12864-019-5608-2

    Figure Lengend Snippet: Comparative qualitative analysis of RNA purified with the Rnaqeuous and Rneasy micro kit. ( a ) Boxplots showing RQN values and ( b ) RNA yield of 10 RNA samples purified from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent RQN values of individual samples. ( c ) Boxplots showing 5′-3′ delta-Cq (dCq) values calculated from a 5′ and a 3′ RT-qPCR assay of 7 RNA samples isolated from 20,000 FACS sorted fli1a:EGFP zebrafish cells with the RNAqueous micro ( left ) and the RNeasy plus micro ( right ) kit. Red dots represent 5′-3′ dCq values of individual samples. * P

    Article Snippet: Based on different quality parameters and comparison of various experimental set-ups, we recommend using the RNeasy Plus Micro Kit for RNA isolation of FACS sorted zebrafish cells.

    Techniques: Purification, FACS, Quantitative RT-PCR, Isolation

    Alteration of keratinocyte gene expression and composition of microbiota of Flg -/- skin. A) RNA sequencing analysis of basal CD49f + keratinocytes (negative for CD45, see also Fig S3), sorted from skin cell suspensions of four 8-12 week-old female Flg -/- mice and 5 littermate controls. Heat map represents expression of genes involved in antigen presentation and the Retnla gene (relative expression, individual read counts corrected for the mean expression of the respective gene), p

    Journal: bioRxiv

    Article Title: Filaggrin deficient mice have a lower threshold for cutaneous allergen sensitization but do not develop spontaneous skin inflammation or atopy

    doi: 10.1101/2020.09.11.293688

    Figure Lengend Snippet: Alteration of keratinocyte gene expression and composition of microbiota of Flg -/- skin. A) RNA sequencing analysis of basal CD49f + keratinocytes (negative for CD45, see also Fig S3), sorted from skin cell suspensions of four 8-12 week-old female Flg -/- mice and 5 littermate controls. Heat map represents expression of genes involved in antigen presentation and the Retnla gene (relative expression, individual read counts corrected for the mean expression of the respective gene), p

    Article Snippet: quantitative RT-PCR Total RNA from sorted CD45-negative cells was isolated using RNeasy® Plus Micro Kit (Qiagen), while RNA from whole ear skin tissue was extracted using RNeasy® Mini Kit (Qiagen), according to the manufacturer’s instructions. cDNA was synthesized using the PrimeScript RT Reagent Kit (Takara).

    Techniques: Expressing, RNA Sequencing Assay, Mouse Assay