rneasy minikit  (Qiagen)


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    Structured Review

    Qiagen rneasy minikit
    Validation of gene silencing in HEp-2C and Vero cells by quantitative PCR (qPCR). siRNAs targeting selected hit genes were transfected into HEp-2C or Vero cells. At 48 h posttransfection, cells were harvested for RNA isolation using a Qiagen <t>RNeasy</t> <t>minikit.</t>
    Rneasy Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 2156 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy minikit/product/Qiagen
    Average 99 stars, based on 2156 article reviews
    Price from $9.99 to $1999.99
    rneasy minikit - by Bioz Stars, 2020-02
    99/100 stars

    Images

    1) Product Images from "Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game"

    Article Title: Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game

    Journal: Journal of Virology

    doi: 10.1128/JVI.01464-15

    Validation of gene silencing in HEp-2C and Vero cells by quantitative PCR (qPCR). siRNAs targeting selected hit genes were transfected into HEp-2C or Vero cells. At 48 h posttransfection, cells were harvested for RNA isolation using a Qiagen RNeasy minikit.
    Figure Legend Snippet: Validation of gene silencing in HEp-2C and Vero cells by quantitative PCR (qPCR). siRNAs targeting selected hit genes were transfected into HEp-2C or Vero cells. At 48 h posttransfection, cells were harvested for RNA isolation using a Qiagen RNeasy minikit.

    Techniques Used: Real-time Polymerase Chain Reaction, Transfection, Isolation

    2) Product Images from "DDX6 (Rck/p54) Is Required for Efficient Hepatitis C Virus Replication but Not for Internal Ribosome Entry Site-Directed Translation ▿"

    Article Title: DDX6 (Rck/p54) Is Required for Efficient Hepatitis C Virus Replication but Not for Internal Ribosome Entry Site-Directed Translation ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.00397-10

    The interaction of DDX6 with HCV core and viral RNA is dependent on the C-terminal domain of DDX6. (A) Schematic representation of EYFP-DDX6 and related mutants, ΔC (C-terminal deletion mutant lacking 183 amino acids), EQ (mutation in the DEAD-box helicase motif II involving the substitution of Glu-247 by Gln), and empty vector expressing EYFP alone. The ΔC mutation removes the second of two conserved RecA domains, identified by the shaded bars within the DDX6 sequence. (B) HJ3-5 HCV-infected FT3-7 cells were transfected with DNA vector expressing various forms of EYFP-DDX6 or just the EYFP. Cell lysates were prepared 2 days later and subjected to coimmunoprecipitation with anti-GFP (rabbit polyclonal; Clontech) antibody. Coimmunoprecipitation samples were immunoblotted with GFP (top panel) and core-specific (bottom panel) antibodies. Input represents 1/20 of the immunoprecipitation (IP) for anti-GFP blot and 1/80 of the IP for HCV core blot. Total RNA isolated from immunoprecipitates using an RNeasy minikit (Qiagen) was subjected to RT-PCR for detection of genomic HCV RNA (primers targeting the NS3 region) and GAPDH mRNA. (C) Huh-7-191/20 cells were induced to express HCV core protein by removing tetracycline from the medium for 3 days. Cells were then transfected with various DNAs and subjected to coimmunoprecipitation exactly as described for panel B.
    Figure Legend Snippet: The interaction of DDX6 with HCV core and viral RNA is dependent on the C-terminal domain of DDX6. (A) Schematic representation of EYFP-DDX6 and related mutants, ΔC (C-terminal deletion mutant lacking 183 amino acids), EQ (mutation in the DEAD-box helicase motif II involving the substitution of Glu-247 by Gln), and empty vector expressing EYFP alone. The ΔC mutation removes the second of two conserved RecA domains, identified by the shaded bars within the DDX6 sequence. (B) HJ3-5 HCV-infected FT3-7 cells were transfected with DNA vector expressing various forms of EYFP-DDX6 or just the EYFP. Cell lysates were prepared 2 days later and subjected to coimmunoprecipitation with anti-GFP (rabbit polyclonal; Clontech) antibody. Coimmunoprecipitation samples were immunoblotted with GFP (top panel) and core-specific (bottom panel) antibodies. Input represents 1/20 of the immunoprecipitation (IP) for anti-GFP blot and 1/80 of the IP for HCV core blot. Total RNA isolated from immunoprecipitates using an RNeasy minikit (Qiagen) was subjected to RT-PCR for detection of genomic HCV RNA (primers targeting the NS3 region) and GAPDH mRNA. (C) Huh-7-191/20 cells were induced to express HCV core protein by removing tetracycline from the medium for 3 days. Cells were then transfected with various DNAs and subjected to coimmunoprecipitation exactly as described for panel B.

    Techniques Used: Mutagenesis, Plasmid Preparation, Expressing, Sequencing, Infection, Transfection, Immunoprecipitation, Isolation, Reverse Transcription Polymerase Chain Reaction

    Related Articles

    Clone Assay:

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    Amplification:

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    Synthesized:

    Article Title: Tramesan, a novel polysaccharide from Trametes versicolor. Structural characterization and biological effects
    Article Snippet: Before RT-PCR analysis, the cells were washed with PBS and total RNA was isolated using RNeasy Minikit (Qiagen). .. Subsequently, cDNA was synthesized using oligo-dT primers and ImProm-IITM reverse transcriptase (Promega) according to the manufacturer’s instructions.

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    Article Snippet: Real-time PCR After homogenization of frozen tissue sections using RNeasy MiniKit® (cat. 74,106, Qiagen, Hilden, Germany) total RNA was extracted, with additional DNase digestion to remove all traces of genomic DNA. .. Total RNA was reverse transcribed into cDNA: cDNA probes were synthesized in 20 μL reaction volume with 1 μg total RNA, 0.5 μg oligo(dT) primer (Promega, Mannheim, Gemany), 40 units of RNasin (Promega, Mannheim, Germany), 0.5 mM dNTP (Biolabs, Frankfurt am Main, Germany), 4 μL 5× transcription buffer and 200 units of Moloney murine leukemia virus (M-MLV) reverse transcriptase (Promega, Mannheim, Germany) for 1 h at 37 °C.

    Article Title: Unexpected Replication Boost by Simeprevir for Simeprevir-Resistant Variants in Genotype 1a Hepatitis C Virus
    Article Snippet: HCV RNA was synthesized with the T7 RiboMAX Express large-scale RNA production system (Promega, Madison, WI) after linearization of the plasmids with XbaI. .. Following treatment with RNase-free DNase to remove the template DNA, RNA was purified by using the RNeasy minikit (Qiagen, Hilden, Germany).

    Quantitative RT-PCR:

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    Real-time Polymerase Chain Reaction:

    Article Title: GRK2 knockdown in mice exacerbates kidney injury and alters renal mechanisms of blood pressure regulation
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    Microarray:

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    Incubation:

    Article Title: Tramesan, a novel polysaccharide from Trametes versicolor. Structural characterization and biological effects
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    Activity Assay:

    Article Title: Broad-Host-Range Expression Reveals Native and Host Regulatory Elements That Influence Heterologous Antibiotic Production in Gram-Negative Bacteria
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    Expressing:

    Article Title: GRK2 knockdown in mice exacerbates kidney injury and alters renal mechanisms of blood pressure regulation
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    Article Title: Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game
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    Article Title: Tramesan, a novel polysaccharide from Trametes versicolor. Structural characterization and biological effects
    Article Snippet: For evaluating gene expression, an amount of B16-F10 stabilized cells (3x105 ) was plated and incubated. .. Before RT-PCR analysis, the cells were washed with PBS and total RNA was isolated using RNeasy Minikit (Qiagen).

    Article Title: Lactobacillus bulgaricus, Lactobacillus rhamnosus and Lactobacillus paracasei Attenuate Salmonella Enteritidis, Salmonella Heidelberg and Salmonella Typhimurium Colonization and Virulence Gene Expression In Vitro
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    Article Title: SHP2 Is Required for BCR-ABL1-Induced Hematologic Neoplasia
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    Article Title: Intracellular interactions of umeclidinium and vilanterol in human airway smooth muscle
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    Hybridization:

    Article Title: NEK7 regulates dendrite morphogenesis in neurons via Eg5-dependent microtubule stabilization
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    Transfection:

    Article Title: Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game
    Article Snippet: .. Forty-eight hours after siRNA transfection, total RNA was isolated from mock-infected and transfected HEp-2C cells and Vero cells using an RNeasy minikit (Qiagen) according to the manufacturer's protocol. .. Expression levels of high-mobility group box 1 protein (HMGB1; a marker for necrosis) and genes of the apoptotic pathway were studied in cells infected with Sabin-1 poliovirus (MOI = 1.0) at 3.5 h prior to RNA isolation.

    Article Title: DDX41 Recognizes RNA/DNA Retroviral Reverse Transcripts and Is Critical for In Vivo Control of Murine Leukemia Virus Infection
    Article Snippet: NR9456 cells, BMDMs, and BMDCs were transfected with the indicated siRNAs ( ) using Lipofectamine RNAiMAX reagent (Invitrogen). .. RNA was isolated using the RNeasy minikit (Qiagen).

    Article Title: Unexpected Replication Boost by Simeprevir for Simeprevir-Resistant Variants in Genotype 1a Hepatitis C Virus
    Article Snippet: Paragraph title: RNA transcription and transfection. ... Following treatment with RNase-free DNase to remove the template DNA, RNA was purified by using the RNeasy minikit (Qiagen, Hilden, Germany).

    Sequencing:

    Article Title: First genome report and analysis of chicken H7N9 influenza viruses with poly-basic amino acids insertion in the hemagglutinin cleavage site
    Article Snippet: Paragraph title: Real time RT-PCR detection and viral genome sequencing ... Viral RNA was extracted using the RNeasy Minikit (Qiagen, Germany).

    Northern Blot:

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    Article Snippet: Paragraph title: Northern blots for HCV RNA. ... Total RNA was isolated from cells by using the RNeasy minikit (Qiagen) and analyzed using reagents supplied with the NorthernMax kit (Applied Biosystems).

    Infection:

    Article Title: Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game
    Article Snippet: Forty-eight hours after siRNA transfection, total RNA was isolated from mock-infected and transfected HEp-2C cells and Vero cells using an RNeasy minikit (Qiagen) according to the manufacturer's protocol. .. Expression levels of high-mobility group box 1 protein (HMGB1; a marker for necrosis) and genes of the apoptotic pathway were studied in cells infected with Sabin-1 poliovirus (MOI = 1.0) at 3.5 h prior to RNA isolation.

    Light Microscopy:

    Article Title: Tramesan, a novel polysaccharide from Trametes versicolor. Structural characterization and biological effects
    Article Snippet: The melanoma cells were then incubated at 37°C for 24 and 48 h and the cell counts were performed with light microscopy at these time points. .. Before RT-PCR analysis, the cells were washed with PBS and total RNA was isolated using RNeasy Minikit (Qiagen).

    Generated:

    Article Title: Tramesan, a novel polysaccharide from Trametes versicolor. Structural characterization and biological effects
    Article Snippet: Melanin content was calculated by interpolating the results with a standard curve, generated by absorbance of known concentrations of synthetic melanin and corrected for the number of cells. .. Before RT-PCR analysis, the cells were washed with PBS and total RNA was isolated using RNeasy Minikit (Qiagen).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game
    Article Snippet: The expression levels of host genes were evaluated by real-time reverse transcription (RT)-PCR using predesigned gene-specific probes and primers (Solaris expression assay; GE Healthcare) and an apoptotic RT2 Profiler PCR array (Qiagen, Germantown, MD) according to the manufacturers' instructions. .. Forty-eight hours after siRNA transfection, total RNA was isolated from mock-infected and transfected HEp-2C cells and Vero cells using an RNeasy minikit (Qiagen) according to the manufacturer's protocol.

    Article Title: Tramesan, a novel polysaccharide from Trametes versicolor. Structural characterization and biological effects
    Article Snippet: .. Before RT-PCR analysis, the cells were washed with PBS and total RNA was isolated using RNeasy Minikit (Qiagen). .. Subsequently, cDNA was synthesized using oligo-dT primers and ImProm-IITM reverse transcriptase (Promega) according to the manufacturer’s instructions.

    Article Title: Glucose Deprivation Induces ATF4-Mediated Apoptosis through TRAIL Death Receptors
    Article Snippet: Paragraph title: RT-PCR analysis of spliced XBP1. ... Total RNA was isolated by using the RNeasy minikit (Qiagen) according to the manufacturer's instructions.

    Article Title: DDX41 Recognizes RNA/DNA Retroviral Reverse Transcripts and Is Critical for In Vivo Control of Murine Leukemia Virus Infection
    Article Snippet: RNA was isolated using the RNeasy minikit (Qiagen). .. All siRNAs used in this study were previously shown to be on target and to decrease both RNA and protein levels ( ). cDNA was made using the SuperScript III first-strand synthesis system for RT-PCR (Invitrogen).

    Article Title: First genome report and analysis of chicken H7N9 influenza viruses with poly-basic amino acids insertion in the hemagglutinin cleavage site
    Article Snippet: Viral RNA was extracted using the RNeasy Minikit (Qiagen, Germany). .. Real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed with the PCR-Fluorescence Detection Kit for H7 Influenza A virus RNA (Cat No. SJ-LG-004–3, Shanghai Biogerm Biological Technology Co., LTD, Shanghai, China) following the manufacturer’s instructions, on the ABI-7500 Real-time PCR system (Applied Biosystems, Foster City, CA).

    Article Title: Renal allograft rejection, lymphocyte infiltration, and de novo donor-specific antibodies in a novel model of non-adherence to immunosuppressive therapy
    Article Snippet: Real-time PCR After homogenization of frozen tissue sections using RNeasy MiniKit® (cat. 74,106, Qiagen, Hilden, Germany) total RNA was extracted, with additional DNase digestion to remove all traces of genomic DNA. .. RT-PCR was performed on ViiA7 detection system in triplicates (Applied Biosystems, Darmstadt, Germany) using QuantiTect SYBR Green PCR Kit (Qiagen, Hilden, Germany).

    Isolation:

    Article Title: Type III Interferon-Mediated Signaling Is Critical for Controlling Live Attenuated Yellow Fever Virus Infection In Vivo
    Article Snippet: .. The isolated tissues (20 to 30 mg) were then resuspended in buffer RLT–1% β-mercaptoethanol (Qiagen, Hilden, Germany), lysed using a TissueLyser (Qiagen, Hilden, Germany) (20 cycles/s for 2 min, 1 min wait, 20 cycles/s for 2 min), and centrifuged at high speed (10,000 rpm) for 10 min. Total RNA was then extracted from the resulting supernatant using an RNeasy minikit (Qiagen, Hilden, Germany) following the manufacturer’s instructions. .. YFV-17D single-step reverse transcription–quantitative real-time PCR.

    Article Title: Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game
    Article Snippet: .. Forty-eight hours after siRNA transfection, total RNA was isolated from mock-infected and transfected HEp-2C cells and Vero cells using an RNeasy minikit (Qiagen) according to the manufacturer's protocol. .. Expression levels of high-mobility group box 1 protein (HMGB1; a marker for necrosis) and genes of the apoptotic pathway were studied in cells infected with Sabin-1 poliovirus (MOI = 1.0) at 3.5 h prior to RNA isolation.

    Article Title: DDX6 (Rck/p54) Is Required for Efficient Hepatitis C Virus Replication but Not for Internal Ribosome Entry Site-Directed Translation ▿
    Article Snippet: .. Total RNA was isolated from cells by using the RNeasy minikit (Qiagen) and analyzed using reagents supplied with the NorthernMax kit (Applied Biosystems). .. Briefly, 5 μg of each RNA sample was resolved on a 0.9% denaturing formaldehyde agarose gel, transferred to a BrightStar-Plus nylon membrane (Applied Biosystems) by downward capillary transfer and hybridized overnight at 68°C with 32 P-labeled antisense riboprobes complementary to the genotype 2a HCV (JFH1) 5′-untranslated region (5′UTR; 340 nucleotides) or β-actin (as a loading control).

    Article Title: Tramesan, a novel polysaccharide from Trametes versicolor. Structural characterization and biological effects
    Article Snippet: .. Before RT-PCR analysis, the cells were washed with PBS and total RNA was isolated using RNeasy Minikit (Qiagen). .. Subsequently, cDNA was synthesized using oligo-dT primers and ImProm-IITM reverse transcriptase (Promega) according to the manufacturer’s instructions.

    Article Title: Lactobacillus bulgaricus, Lactobacillus rhamnosus and Lactobacillus paracasei Attenuate Salmonella Enteritidis, Salmonella Heidelberg and Salmonella Typhimurium Colonization and Virulence Gene Expression In Vitro
    Article Snippet: .. RNA was isolated using RNeasy minikit (Qiagen, Germantown, MD, USA) according to the manufacturer’s protocol. .. RNA quantification was performed using the Nanodrop (Eppendorf, Enfield, CT, USA), and cDNA was synthesized using the iScript reverse transcriptase kit (Biorad, Hercules, CA, USA). cDNA was used as a template for the Salmonella virulence gene expression assay.

    Article Title: Glucose Deprivation Induces ATF4-Mediated Apoptosis through TRAIL Death Receptors
    Article Snippet: .. Total RNA was isolated by using the RNeasy minikit (Qiagen) according to the manufacturer's instructions. .. One microgram of RNA under each condition was retrotranscribed at cDNA by using the High-Capacity cDNA reverse transcription (RT) kit from Applied Biosystems.

    Article Title: DDX41 Recognizes RNA/DNA Retroviral Reverse Transcripts and Is Critical for In Vivo Control of Murine Leukemia Virus Infection
    Article Snippet: .. RNA was isolated using the RNeasy minikit (Qiagen). .. All siRNAs used in this study were previously shown to be on target and to decrease both RNA and protein levels ( ). cDNA was made using the SuperScript III first-strand synthesis system for RT-PCR (Invitrogen).

    Article Title: Broad-Host-Range Expression Reveals Native and Host Regulatory Elements That Influence Heterologous Antibiotic Production in Gram-Negative Bacteria
    Article Snippet: .. RNA was isolated with an RNeasy minikit (Qiagen), and cell lysis was obtained by homogenization with the FastPrep system (BIO 101 Inc.) (five cycles of 30 s at 5.5 speed). .. Turbo DNase (Thermo Fisher Scientific) treatment was carried out for 2 h in accordance with the manufacturer’s instructions.

    Article Title: Intracellular interactions of umeclidinium and vilanterol in human airway smooth muscle
    Article Snippet: .. Total RNA was isolated from cells using an RNeasy minikit (Qiagen, Crawley, UK) and converted to complementary DNA using random primers and avian myeloblastosis virus reverse transcriptase (Pro-mega, Southampton, UK). .. Messenger RNA (mRNA) expression was determined by real-time quantitative polymerase chain reaction (Rotor-Gene 3000; Corbett Research, St Neots, UK) using SYBR Green PCR Mix Reagent (Qiagen) and normalized to 18S RNA expression.

    Labeling:

    Article Title: NEK7 regulates dendrite morphogenesis in neurons via Eg5-dependent microtubule stabilization
    Article Snippet: Microarray and qRT-PCR For two independent microarray replicas, total RNA was extracted from mouse embryo hippocampal neuron cultures at different time points of differentiation (0 DIV, 20 h, 3 DIV, 6 DIV, 12 DIV, 15 DIV) using a commercial RNeasy minikit (Qiagen) according to the manufacturer’s instructions. .. Then, 25 ng total RNA was amplified using the TransPlex® Complete Whole Transcriptome Amplification Kit (Sigma; reference WTA2) and subsequently labeled using GeneChip Mapping 10K Xba Assay Kit (Affymetrix; catalog no. 900441), according to the manufacturer’s instructions.

    Mouse Assay:

    Article Title: Type III Interferon-Mediated Signaling Is Critical for Controlling Live Attenuated Yellow Fever Virus Infection In Vivo
    Article Snippet: At the indicated endpoints, mice were euthanized via exsanguination under ketamine/xylazine anesthesia. .. The isolated tissues (20 to 30 mg) were then resuspended in buffer RLT–1% β-mercaptoethanol (Qiagen, Hilden, Germany), lysed using a TissueLyser (Qiagen, Hilden, Germany) (20 cycles/s for 2 min, 1 min wait, 20 cycles/s for 2 min), and centrifuged at high speed (10,000 rpm) for 10 min. Total RNA was then extracted from the resulting supernatant using an RNeasy minikit (Qiagen, Hilden, Germany) following the manufacturer’s instructions.

    Polymerase Chain Reaction:

    Article Title: Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game
    Article Snippet: The expression levels of host genes were evaluated by real-time reverse transcription (RT)-PCR using predesigned gene-specific probes and primers (Solaris expression assay; GE Healthcare) and an apoptotic RT2 Profiler PCR array (Qiagen, Germantown, MD) according to the manufacturers' instructions. .. Forty-eight hours after siRNA transfection, total RNA was isolated from mock-infected and transfected HEp-2C cells and Vero cells using an RNeasy minikit (Qiagen) according to the manufacturer's protocol.

    Article Title: Tramesan, a novel polysaccharide from Trametes versicolor. Structural characterization and biological effects
    Article Snippet: Before RT-PCR analysis, the cells were washed with PBS and total RNA was isolated using RNeasy Minikit (Qiagen). .. RT-PCR was carried out in 15 μL (total volume) with SYBR green PCR Master Mix (Bio-Rad) and 200 nM of each primer.

    Article Title: DDX41 Recognizes RNA/DNA Retroviral Reverse Transcripts and Is Critical for In Vivo Control of Murine Leukemia Virus Infection
    Article Snippet: RNA was isolated using the RNeasy minikit (Qiagen). .. RT-PCR was performed using the Power SYBR green PCR master mix kit (Applied Biosystems).

    Article Title: First genome report and analysis of chicken H7N9 influenza viruses with poly-basic amino acids insertion in the hemagglutinin cleavage site
    Article Snippet: Viral RNA was extracted using the RNeasy Minikit (Qiagen, Germany). .. Real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed with the PCR-Fluorescence Detection Kit for H7 Influenza A virus RNA (Cat No. SJ-LG-004–3, Shanghai Biogerm Biological Technology Co., LTD, Shanghai, China) following the manufacturer’s instructions, on the ABI-7500 Real-time PCR system (Applied Biosystems, Foster City, CA).

    Article Title: Renal allograft rejection, lymphocyte infiltration, and de novo donor-specific antibodies in a novel model of non-adherence to immunosuppressive therapy
    Article Snippet: Real-time PCR After homogenization of frozen tissue sections using RNeasy MiniKit® (cat. 74,106, Qiagen, Hilden, Germany) total RNA was extracted, with additional DNase digestion to remove all traces of genomic DNA. .. RT-PCR was performed on ViiA7 detection system in triplicates (Applied Biosystems, Darmstadt, Germany) using QuantiTect SYBR Green PCR Kit (Qiagen, Hilden, Germany).

    Article Title: Broad-Host-Range Expression Reveals Native and Host Regulatory Elements That Influence Heterologous Antibiotic Production in Gram-Negative Bacteria
    Article Snippet: RNA was isolated with an RNeasy minikit (Qiagen), and cell lysis was obtained by homogenization with the FastPrep system (BIO 101 Inc.) (five cycles of 30 s at 5.5 speed). .. The RNA was checked on an agarose gel, and the absence of genomic DNA in the sample was confirmed by PCR with primers for the housekeeping gene cysG ( ). cDNA synthesis was carried out with SuperScript IV reverse transcriptase (Thermo Fisher Scientific), 50-pg/μl random hexamers, and 500 ng of RNA in accordance with the manufacturer’s protocol.

    Article Title: Intracellular interactions of umeclidinium and vilanterol in human airway smooth muscle
    Article Snippet: Total RNA was isolated from cells using an RNeasy minikit (Qiagen, Crawley, UK) and converted to complementary DNA using random primers and avian myeloblastosis virus reverse transcriptase (Pro-mega, Southampton, UK). .. Messenger RNA (mRNA) expression was determined by real-time quantitative polymerase chain reaction (Rotor-Gene 3000; Corbett Research, St Neots, UK) using SYBR Green PCR Mix Reagent (Qiagen) and normalized to 18S RNA expression.

    Staining:

    Article Title: NEK7 regulates dendrite morphogenesis in neurons via Eg5-dependent microtubule stabilization
    Article Snippet: Microarray and qRT-PCR For two independent microarray replicas, total RNA was extracted from mouse embryo hippocampal neuron cultures at different time points of differentiation (0 DIV, 20 h, 3 DIV, 6 DIV, 12 DIV, 15 DIV) using a commercial RNeasy minikit (Qiagen) according to the manufacturer’s instructions. .. Next, 8 µg of complementary DNA (cDNA) was hybridized per microarray at Mouse Gene 1.0 ST for 16 h of hybridization at 45 °C, and washing and staining of microarrays was performed using a Fluidics Station 450 (Affymetrix, Santa Clara, CA).

    Purification:

    Article Title: Unexpected Replication Boost by Simeprevir for Simeprevir-Resistant Variants in Genotype 1a Hepatitis C Virus
    Article Snippet: .. Following treatment with RNase-free DNase to remove the template DNA, RNA was purified by using the RNeasy minikit (Qiagen, Hilden, Germany). .. RNA transfection was carried out with the TransIT mRNA transfection kit (Mirus Bio LLC, Madison, WI), according to the manufacturer's protocol.

    Software:

    Article Title: DDX6 (Rck/p54) Is Required for Efficient Hepatitis C Virus Replication but Not for Internal Ribosome Entry Site-Directed Translation ▿
    Article Snippet: Total RNA was isolated from cells by using the RNeasy minikit (Qiagen) and analyzed using reagents supplied with the NorthernMax kit (Applied Biosystems). .. After extensive washing, the membranes were scanned on a Personal Molecular Imager (Bio-Rad), and the band densities quantified with Quantity One software (Bio-Rad).

    SYBR Green Assay:

    Article Title: Tramesan, a novel polysaccharide from Trametes versicolor. Structural characterization and biological effects
    Article Snippet: Before RT-PCR analysis, the cells were washed with PBS and total RNA was isolated using RNeasy Minikit (Qiagen). .. RT-PCR was carried out in 15 μL (total volume) with SYBR green PCR Master Mix (Bio-Rad) and 200 nM of each primer.

    Article Title: DDX41 Recognizes RNA/DNA Retroviral Reverse Transcripts and Is Critical for In Vivo Control of Murine Leukemia Virus Infection
    Article Snippet: RNA was isolated using the RNeasy minikit (Qiagen). .. RT-PCR was performed using the Power SYBR green PCR master mix kit (Applied Biosystems).

    Article Title: Renal allograft rejection, lymphocyte infiltration, and de novo donor-specific antibodies in a novel model of non-adherence to immunosuppressive therapy
    Article Snippet: Real-time PCR After homogenization of frozen tissue sections using RNeasy MiniKit® (cat. 74,106, Qiagen, Hilden, Germany) total RNA was extracted, with additional DNase digestion to remove all traces of genomic DNA. .. RT-PCR was performed on ViiA7 detection system in triplicates (Applied Biosystems, Darmstadt, Germany) using QuantiTect SYBR Green PCR Kit (Qiagen, Hilden, Germany).

    Article Title: Intracellular interactions of umeclidinium and vilanterol in human airway smooth muscle
    Article Snippet: Total RNA was isolated from cells using an RNeasy minikit (Qiagen, Crawley, UK) and converted to complementary DNA using random primers and avian myeloblastosis virus reverse transcriptase (Pro-mega, Southampton, UK). .. Messenger RNA (mRNA) expression was determined by real-time quantitative polymerase chain reaction (Rotor-Gene 3000; Corbett Research, St Neots, UK) using SYBR Green PCR Mix Reagent (Qiagen) and normalized to 18S RNA expression.

    RNA Extraction:

    Article Title: Type III Interferon-Mediated Signaling Is Critical for Controlling Live Attenuated Yellow Fever Virus Infection In Vivo
    Article Snippet: Paragraph title: RNA extraction from serum and tissues. ... The isolated tissues (20 to 30 mg) were then resuspended in buffer RLT–1% β-mercaptoethanol (Qiagen, Hilden, Germany), lysed using a TissueLyser (Qiagen, Hilden, Germany) (20 cycles/s for 2 min, 1 min wait, 20 cycles/s for 2 min), and centrifuged at high speed (10,000 rpm) for 10 min. Total RNA was then extracted from the resulting supernatant using an RNeasy minikit (Qiagen, Hilden, Germany) following the manufacturer’s instructions.

    Article Title: GRK2 knockdown in mice exacerbates kidney injury and alters renal mechanisms of blood pressure regulation
    Article Snippet: Paragraph title: RNA extraction and quantitative Real-Time Polymerase Chain Reaction ... Total RNA was obtained by using the RNeasy Minikit (Qiagen, Valencia, MD, USA) according manufacturer’s instructions.

    RNA Expression:

    Article Title: Intracellular interactions of umeclidinium and vilanterol in human airway smooth muscle
    Article Snippet: Total RNA was isolated from cells using an RNeasy minikit (Qiagen, Crawley, UK) and converted to complementary DNA using random primers and avian myeloblastosis virus reverse transcriptase (Pro-mega, Southampton, UK). .. Messenger RNA (mRNA) expression was determined by real-time quantitative polymerase chain reaction (Rotor-Gene 3000; Corbett Research, St Neots, UK) using SYBR Green PCR Mix Reagent (Qiagen) and normalized to 18S RNA expression.

    Agarose Gel Electrophoresis:

    Article Title: DDX6 (Rck/p54) Is Required for Efficient Hepatitis C Virus Replication but Not for Internal Ribosome Entry Site-Directed Translation ▿
    Article Snippet: Total RNA was isolated from cells by using the RNeasy minikit (Qiagen) and analyzed using reagents supplied with the NorthernMax kit (Applied Biosystems). .. Briefly, 5 μg of each RNA sample was resolved on a 0.9% denaturing formaldehyde agarose gel, transferred to a BrightStar-Plus nylon membrane (Applied Biosystems) by downward capillary transfer and hybridized overnight at 68°C with 32 P-labeled antisense riboprobes complementary to the genotype 2a HCV (JFH1) 5′-untranslated region (5′UTR; 340 nucleotides) or β-actin (as a loading control).

    Article Title: Broad-Host-Range Expression Reveals Native and Host Regulatory Elements That Influence Heterologous Antibiotic Production in Gram-Negative Bacteria
    Article Snippet: RNA was isolated with an RNeasy minikit (Qiagen), and cell lysis was obtained by homogenization with the FastPrep system (BIO 101 Inc.) (five cycles of 30 s at 5.5 speed). .. The RNA was checked on an agarose gel, and the absence of genomic DNA in the sample was confirmed by PCR with primers for the housekeeping gene cysG ( ). cDNA synthesis was carried out with SuperScript IV reverse transcriptase (Thermo Fisher Scientific), 50-pg/μl random hexamers, and 500 ng of RNA in accordance with the manufacturer’s protocol.

    Homogenization:

    Article Title: Renal allograft rejection, lymphocyte infiltration, and de novo donor-specific antibodies in a novel model of non-adherence to immunosuppressive therapy
    Article Snippet: .. Real-time PCR After homogenization of frozen tissue sections using RNeasy MiniKit® (cat. 74,106, Qiagen, Hilden, Germany) total RNA was extracted, with additional DNase digestion to remove all traces of genomic DNA. .. Total RNA was reverse transcribed into cDNA: cDNA probes were synthesized in 20 μL reaction volume with 1 μg total RNA, 0.5 μg oligo(dT) primer (Promega, Mannheim, Gemany), 40 units of RNasin (Promega, Mannheim, Germany), 0.5 mM dNTP (Biolabs, Frankfurt am Main, Germany), 4 μL 5× transcription buffer and 200 units of Moloney murine leukemia virus (M-MLV) reverse transcriptase (Promega, Mannheim, Germany) for 1 h at 37 °C.

    Article Title: Broad-Host-Range Expression Reveals Native and Host Regulatory Elements That Influence Heterologous Antibiotic Production in Gram-Negative Bacteria
    Article Snippet: .. RNA was isolated with an RNeasy minikit (Qiagen), and cell lysis was obtained by homogenization with the FastPrep system (BIO 101 Inc.) (five cycles of 30 s at 5.5 speed). .. Turbo DNase (Thermo Fisher Scientific) treatment was carried out for 2 h in accordance with the manufacturer’s instructions.

    Marker:

    Article Title: Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game
    Article Snippet: Forty-eight hours after siRNA transfection, total RNA was isolated from mock-infected and transfected HEp-2C cells and Vero cells using an RNeasy minikit (Qiagen) according to the manufacturer's protocol. .. Expression levels of high-mobility group box 1 protein (HMGB1; a marker for necrosis) and genes of the apoptotic pathway were studied in cells infected with Sabin-1 poliovirus (MOI = 1.0) at 3.5 h prior to RNA isolation.

    Lysis:

    Article Title: Broad-Host-Range Expression Reveals Native and Host Regulatory Elements That Influence Heterologous Antibiotic Production in Gram-Negative Bacteria
    Article Snippet: .. RNA was isolated with an RNeasy minikit (Qiagen), and cell lysis was obtained by homogenization with the FastPrep system (BIO 101 Inc.) (five cycles of 30 s at 5.5 speed). .. Turbo DNase (Thermo Fisher Scientific) treatment was carried out for 2 h in accordance with the manufacturer’s instructions.

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  • 95
    Qiagen rneasy plant minikit
    Autoradiogram of 5% PAGE analysis of self-cleavage experiments using either 2′,5′- or 3′,5′-C-PLMVd. Radioactive transcripts (C-PLMVd) were added to the ground leaf powder, and the RNA was extracted using the <t>RNeasy</t> plant <t>minikit</t> and incubated under self-cleavage conditions with or without prior heat denaturation and snap-cooling treatments. Lanes 1 and 6, untreated L-PLMVd (control); lanes 2 to 5 and 7 to 10, 3′,5′- and 2′,5′-C-PLMVd, respectively; lanes 2 and 7, C-PLMVd extracted and incubated under self-cleavage conditions without heat denaturation and snap-cooling steps; lanes 3 and 8, like lanes 2 and 7, except that the extraction was performed in the presence of additional 5 mM EDTA in all buffers; lanes 4 and 9, like lanes 2 and 7, except that the samples were heat denatured and snap-cooled prior to incubation under self-cleavage conditions; lanes 5 and 10, like lanes 4 and 9, except that the extraction was performed in the presence of additional 5 mM EDTA. Adjacent to the gel, the positions of the C-PLMVd and L-PLMVd transcripts are used as size references.
    Rneasy Plant Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 95/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy plant minikit/product/Qiagen
    Average 95 stars, based on 69 article reviews
    Price from $9.99 to $1999.99
    rneasy plant minikit - by Bioz Stars, 2020-02
    95/100 stars
      Buy from Supplier

    99
    Qiagen rneasy minikit
    Validation of gene silencing in HEp-2C and Vero cells by quantitative PCR (qPCR). siRNAs targeting selected hit genes were transfected into HEp-2C or Vero cells. At 48 h posttransfection, cells were harvested for RNA isolation using a Qiagen <t>RNeasy</t> <t>minikit.</t>
    Rneasy Minikit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 2288 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rneasy minikit/product/Qiagen
    Average 99 stars, based on 2288 article reviews
    Price from $9.99 to $1999.99
    rneasy minikit - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    Image Search Results


    Autoradiogram of 5% PAGE analysis of self-cleavage experiments using either 2′,5′- or 3′,5′-C-PLMVd. Radioactive transcripts (C-PLMVd) were added to the ground leaf powder, and the RNA was extracted using the RNeasy plant minikit and incubated under self-cleavage conditions with or without prior heat denaturation and snap-cooling treatments. Lanes 1 and 6, untreated L-PLMVd (control); lanes 2 to 5 and 7 to 10, 3′,5′- and 2′,5′-C-PLMVd, respectively; lanes 2 and 7, C-PLMVd extracted and incubated under self-cleavage conditions without heat denaturation and snap-cooling steps; lanes 3 and 8, like lanes 2 and 7, except that the extraction was performed in the presence of additional 5 mM EDTA in all buffers; lanes 4 and 9, like lanes 2 and 7, except that the samples were heat denatured and snap-cooled prior to incubation under self-cleavage conditions; lanes 5 and 10, like lanes 4 and 9, except that the extraction was performed in the presence of additional 5 mM EDTA. Adjacent to the gel, the positions of the C-PLMVd and L-PLMVd transcripts are used as size references.

    Journal: Journal of Virology

    Article Title: Natural 2?,5?-Phosphodiester Bonds Found at the Ligation Sites of Peach Latent Mosaic Viroid

    doi: 10.1128/JVI.75.1.19-25.2001

    Figure Lengend Snippet: Autoradiogram of 5% PAGE analysis of self-cleavage experiments using either 2′,5′- or 3′,5′-C-PLMVd. Radioactive transcripts (C-PLMVd) were added to the ground leaf powder, and the RNA was extracted using the RNeasy plant minikit and incubated under self-cleavage conditions with or without prior heat denaturation and snap-cooling treatments. Lanes 1 and 6, untreated L-PLMVd (control); lanes 2 to 5 and 7 to 10, 3′,5′- and 2′,5′-C-PLMVd, respectively; lanes 2 and 7, C-PLMVd extracted and incubated under self-cleavage conditions without heat denaturation and snap-cooling steps; lanes 3 and 8, like lanes 2 and 7, except that the extraction was performed in the presence of additional 5 mM EDTA in all buffers; lanes 4 and 9, like lanes 2 and 7, except that the samples were heat denatured and snap-cooled prior to incubation under self-cleavage conditions; lanes 5 and 10, like lanes 4 and 9, except that the extraction was performed in the presence of additional 5 mM EDTA. Adjacent to the gel, the positions of the C-PLMVd and L-PLMVd transcripts are used as size references.

    Article Snippet: The first procedure used an RNeasy plant minikit (Qiagen) to prepare RNA from 50 to 100 mg of tissue as specified by the manufacturer.

    Techniques: Polyacrylamide Gel Electrophoresis, Incubation

    Validation of gene silencing in HEp-2C and Vero cells by quantitative PCR (qPCR). siRNAs targeting selected hit genes were transfected into HEp-2C or Vero cells. At 48 h posttransfection, cells were harvested for RNA isolation using a Qiagen RNeasy minikit.

    Journal: Journal of Virology

    Article Title: Engineering Enhanced Vaccine Cell Lines To Eradicate Vaccine-Preventable Diseases: the Polio End Game

    doi: 10.1128/JVI.01464-15

    Figure Lengend Snippet: Validation of gene silencing in HEp-2C and Vero cells by quantitative PCR (qPCR). siRNAs targeting selected hit genes were transfected into HEp-2C or Vero cells. At 48 h posttransfection, cells were harvested for RNA isolation using a Qiagen RNeasy minikit.

    Article Snippet: Forty-eight hours after siRNA transfection, total RNA was isolated from mock-infected and transfected HEp-2C cells and Vero cells using an RNeasy minikit (Qiagen) according to the manufacturer's protocol.

    Techniques: Real-time Polymerase Chain Reaction, Transfection, Isolation

    The interaction of DDX6 with HCV core and viral RNA is dependent on the C-terminal domain of DDX6. (A) Schematic representation of EYFP-DDX6 and related mutants, ΔC (C-terminal deletion mutant lacking 183 amino acids), EQ (mutation in the DEAD-box helicase motif II involving the substitution of Glu-247 by Gln), and empty vector expressing EYFP alone. The ΔC mutation removes the second of two conserved RecA domains, identified by the shaded bars within the DDX6 sequence. (B) HJ3-5 HCV-infected FT3-7 cells were transfected with DNA vector expressing various forms of EYFP-DDX6 or just the EYFP. Cell lysates were prepared 2 days later and subjected to coimmunoprecipitation with anti-GFP (rabbit polyclonal; Clontech) antibody. Coimmunoprecipitation samples were immunoblotted with GFP (top panel) and core-specific (bottom panel) antibodies. Input represents 1/20 of the immunoprecipitation (IP) for anti-GFP blot and 1/80 of the IP for HCV core blot. Total RNA isolated from immunoprecipitates using an RNeasy minikit (Qiagen) was subjected to RT-PCR for detection of genomic HCV RNA (primers targeting the NS3 region) and GAPDH mRNA. (C) Huh-7-191/20 cells were induced to express HCV core protein by removing tetracycline from the medium for 3 days. Cells were then transfected with various DNAs and subjected to coimmunoprecipitation exactly as described for panel B.

    Journal: Journal of Virology

    Article Title: DDX6 (Rck/p54) Is Required for Efficient Hepatitis C Virus Replication but Not for Internal Ribosome Entry Site-Directed Translation ▿

    doi: 10.1128/JVI.00397-10

    Figure Lengend Snippet: The interaction of DDX6 with HCV core and viral RNA is dependent on the C-terminal domain of DDX6. (A) Schematic representation of EYFP-DDX6 and related mutants, ΔC (C-terminal deletion mutant lacking 183 amino acids), EQ (mutation in the DEAD-box helicase motif II involving the substitution of Glu-247 by Gln), and empty vector expressing EYFP alone. The ΔC mutation removes the second of two conserved RecA domains, identified by the shaded bars within the DDX6 sequence. (B) HJ3-5 HCV-infected FT3-7 cells were transfected with DNA vector expressing various forms of EYFP-DDX6 or just the EYFP. Cell lysates were prepared 2 days later and subjected to coimmunoprecipitation with anti-GFP (rabbit polyclonal; Clontech) antibody. Coimmunoprecipitation samples were immunoblotted with GFP (top panel) and core-specific (bottom panel) antibodies. Input represents 1/20 of the immunoprecipitation (IP) for anti-GFP blot and 1/80 of the IP for HCV core blot. Total RNA isolated from immunoprecipitates using an RNeasy minikit (Qiagen) was subjected to RT-PCR for detection of genomic HCV RNA (primers targeting the NS3 region) and GAPDH mRNA. (C) Huh-7-191/20 cells were induced to express HCV core protein by removing tetracycline from the medium for 3 days. Cells were then transfected with various DNAs and subjected to coimmunoprecipitation exactly as described for panel B.

    Article Snippet: Total RNA was isolated from cells by using the RNeasy minikit (Qiagen) and analyzed using reagents supplied with the NorthernMax kit (Applied Biosystems).

    Techniques: Mutagenesis, Plasmid Preparation, Expressing, Sequencing, Infection, Transfection, Immunoprecipitation, Isolation, Reverse Transcription Polymerase Chain Reaction