qrt pcr assay rneasy lipid tissue mini kit  (Qiagen)


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    RNeasy Lipid Tissue Mini Kit
    Description:
    For purification of up to 100 µg total RNA from fatty tissues and other types of tissue Kit contents Qiagen RNeasy Lipid Tissue Mini Kit 50 preps 10 to 100mg Sample 30 to 100L Elution Volume Fatty Tissue Sample RNA Purification 2 4 to 10mg Yield Spin Column Format Silica Technology 45 min Time Run Ideal for PCR Real time PCR Microarray Includes 50 RNeasy Mini Spin Columns Collection Tubes 1 5 and 2mL QIAzol Lysis Reagent RNase free Reagents and Buffers Benefits Optimized lysis conditions for fatty tissues and other tissues High yields of total RNA without phenol contamination High quality RNA for all downstream applicatio
    Catalog Number:
    74804
    Price:
    401
    Category:
    RNeasy Lipid Tissue Mini Kit
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    Structured Review

    Qiagen qrt pcr assay rneasy lipid tissue mini kit
    RNeasy Lipid Tissue Mini Kit
    For purification of up to 100 µg total RNA from fatty tissues and other types of tissue Kit contents Qiagen RNeasy Lipid Tissue Mini Kit 50 preps 10 to 100mg Sample 30 to 100L Elution Volume Fatty Tissue Sample RNA Purification 2 4 to 10mg Yield Spin Column Format Silica Technology 45 min Time Run Ideal for PCR Real time PCR Microarray Includes 50 RNeasy Mini Spin Columns Collection Tubes 1 5 and 2mL QIAzol Lysis Reagent RNase free Reagents and Buffers Benefits Optimized lysis conditions for fatty tissues and other tissues High yields of total RNA without phenol contamination High quality RNA for all downstream applicatio
    https://www.bioz.com/result/qrt pcr assay rneasy lipid tissue mini kit/product/Qiagen
    Average 95 stars, based on 929 article reviews
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    qrt pcr assay rneasy lipid tissue mini kit - by Bioz Stars, 2020-02
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    Images

    1) Product Images from "Phytochemical analysis and antioxidant and anticancer activities of mastic gum resin from Pistacia atlantica subspecies kurdica"

    Article Title: Phytochemical analysis and antioxidant and anticancer activities of mastic gum resin from Pistacia atlantica subspecies kurdica

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S170827

    mRNA expression levels of Bcl-2 , Bax , and Cyt-c normalized to the transcription levels of β-actin and GAPDH . The qRT-PCR analysis was performed on COLO205 cells treated with 5.2 ± 0.8 μg/mL MGR. The experiment was done in triplicates, and data are expressed as mean ± SD. *Significant difference from control ( P
    Figure Legend Snippet: mRNA expression levels of Bcl-2 , Bax , and Cyt-c normalized to the transcription levels of β-actin and GAPDH . The qRT-PCR analysis was performed on COLO205 cells treated with 5.2 ± 0.8 μg/mL MGR. The experiment was done in triplicates, and data are expressed as mean ± SD. *Significant difference from control ( P

    Techniques Used: Expressing, Quantitative RT-PCR

    2) Product Images from "Egr-1 Regulates Expression of the Glial Scar Component Phosphacan in Astrocytes after Experimental Stroke"

    Article Title: Egr-1 Regulates Expression of the Glial Scar Component Phosphacan in Astrocytes after Experimental Stroke

    Journal: The American Journal of Pathology

    doi: 10.2353/ajpath.2008.070648

    Phosphacan accumulation is impaired in the glial scar of Egr-1-deficient mice. A and B: Measurement of GFAP ( A ) and phosphacan ( B ) RNA expression levels 10 days after permanent occlusion of the MCA. RNA samples were isolated from wild-type (+/+) and Egr-1-deficient (−/−) mice and from the infarcted, left brain (L) and contralateral, right brain (R) hemispheres. Results were normalized to β-actin RNA expression levels setting arbitrarily the value of one wild-type sample from the right hemisphere as 100. Statistical analysis was performed with the Mann-Whitney rank sum test. Wild-type and Egr-1-deficient mice show comparable GFAP induction. Egr-1 −/− mice show a statistically significant reduction of phosphacan levels in the infarcted hemisphere compared with the contralateral site ( P = 0.001). The expression levels of phosphacan are also statistically significant different between the infarcted hemispheres of Egr-1 −/− mice and their wild-type littermates ( P ≤ 0.001). In contrast, there is no statistically significant change of phosphacan expression between both hemispheres ( P = 0.163) in wild-type mice and between noninfarcted hemispheres of WT and KO mice ( P = 0.380). C: Western blot analysis of brain tissue isolated from the glial scar 10 days after occlusion of the MCA and control (c) tissue from the contralateral brain hemisphere. Quantitative image analysis demonstrates that GFAP up-regulation after stroke is comparable in Egr-1 −/− and wild-type brain tissue, ie, 4.4 times in wild-type samples (SD, 0.6) and 4.7 times in Egr-1 −/− mice (SD, 1.8). The levels of the two phosphacan isoforms detected by the KAF13 antibody, namely the secreted form of phosphacan (Pcan) as well as the transmembrane isoform receptor protein tyrosine phosphatase β (RPTPβs) are up-regulated in the glial scar of wild types by 1.49-fold (SD, 0.14), but not in the glial scar of Egr-1-deficient animals (0.89-fold; SD, 0.26). β-Tubulin serves as loading control. Protein sizes are indicated in kDa.
    Figure Legend Snippet: Phosphacan accumulation is impaired in the glial scar of Egr-1-deficient mice. A and B: Measurement of GFAP ( A ) and phosphacan ( B ) RNA expression levels 10 days after permanent occlusion of the MCA. RNA samples were isolated from wild-type (+/+) and Egr-1-deficient (−/−) mice and from the infarcted, left brain (L) and contralateral, right brain (R) hemispheres. Results were normalized to β-actin RNA expression levels setting arbitrarily the value of one wild-type sample from the right hemisphere as 100. Statistical analysis was performed with the Mann-Whitney rank sum test. Wild-type and Egr-1-deficient mice show comparable GFAP induction. Egr-1 −/− mice show a statistically significant reduction of phosphacan levels in the infarcted hemisphere compared with the contralateral site ( P = 0.001). The expression levels of phosphacan are also statistically significant different between the infarcted hemispheres of Egr-1 −/− mice and their wild-type littermates ( P ≤ 0.001). In contrast, there is no statistically significant change of phosphacan expression between both hemispheres ( P = 0.163) in wild-type mice and between noninfarcted hemispheres of WT and KO mice ( P = 0.380). C: Western blot analysis of brain tissue isolated from the glial scar 10 days after occlusion of the MCA and control (c) tissue from the contralateral brain hemisphere. Quantitative image analysis demonstrates that GFAP up-regulation after stroke is comparable in Egr-1 −/− and wild-type brain tissue, ie, 4.4 times in wild-type samples (SD, 0.6) and 4.7 times in Egr-1 −/− mice (SD, 1.8). The levels of the two phosphacan isoforms detected by the KAF13 antibody, namely the secreted form of phosphacan (Pcan) as well as the transmembrane isoform receptor protein tyrosine phosphatase β (RPTPβs) are up-regulated in the glial scar of wild types by 1.49-fold (SD, 0.14), but not in the glial scar of Egr-1-deficient animals (0.89-fold; SD, 0.26). β-Tubulin serves as loading control. Protein sizes are indicated in kDa.

    Techniques Used: Mouse Assay, RNA Expression, Isolation, MANN-WHITNEY, Expressing, Western Blot

    3) Product Images from "Gene Expression in the Spinal Cord in Female Lewis Rats with Experimental Autoimmune Encephalomyelitis Induced with Myelin Basic Protein"

    Article Title: Gene Expression in the Spinal Cord in Female Lewis Rats with Experimental Autoimmune Encephalomyelitis Induced with Myelin Basic Protein

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0048555

    RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.
    Figure Legend Snippet: RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Electrophoresis, Amplification, Expressing, Microarray, Derivative Assay

    4) Product Images from "Gene Expression in the Spinal Cord in Female Lewis Rats with Experimental Autoimmune Encephalomyelitis Induced with Myelin Basic Protein"

    Article Title: Gene Expression in the Spinal Cord in Female Lewis Rats with Experimental Autoimmune Encephalomyelitis Induced with Myelin Basic Protein

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0048555

    RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.
    Figure Legend Snippet: RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Electrophoresis, Amplification, Expressing, Microarray, Derivative Assay

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    Synthesized:

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    Quantitative RT-PCR:

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    Real-time Polymerase Chain Reaction:

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    Microarray:

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    Quantitation Assay:

    Article Title: Perturbations of Fibroblast Growth Factors 19 and 21 in Type 2 Diabetes
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    Expressing:

    Article Title: Perturbations of Fibroblast Growth Factors 19 and 21 in Type 2 Diabetes
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    Article Title: Secretion of SDF-1α by bone marrow-derived stromal cells enhances skin wound healing of C57BL/6 mice exposed to ionizing radiation
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    Article Title: Effects of internal low-dose irradiation from 131I on gene expression in normal tissues in Balb/c mice
    Article Snippet: Paragraph title: Gene expression analysis ... Total RNA was extracted using the RNeasy Lipid Tissue Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions.

    Hybridization:

    Article Title: Obesity and prostate cancer: gene expression signature of human periprostatic adipose tissue
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    SYBR Green Assay:

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    Article Title: Sustained Exendin-4 Secretion through Gene Therapy Targeting Salivary Glands in Two Different Rodent Models of Obesity/Type 2 Diabetes
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    Polymerase Chain Reaction:

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    Article Title: Regulation of energy metabolism during social interactions in rainbow trout: a role for AMP-activated protein kinase
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    Article Title: Selected imprinting of INS in the marsupial
    Article Snippet: RNA and DNA extraction and RT-PCR Individual genotypes were identified using PCR and direct sequencing of genomic DNA extracted from approximately 20 mg snap-frozen tissue using a Wizard Genomic DNA Purification Kit (Promega, Madison, WI, USA). .. Total RNA was extracted from mammary glands using the RNeasy Lipid Tissue Mini Kit (Qiagen,Hilden, Germany) or from other tissues using Tri-Reagent (Ambion, Austin, Texas, USA) as described by the manufacturer with a final elution of RNA in 60 to 80 μl RNAsecure H2 O (a dilution of 1/24 μl RNAsecure to water;Ambion).

    Article Title: Secretion of SDF-1α by bone marrow-derived stromal cells enhances skin wound healing of C57BL/6 mice exposed to ionizing radiation
    Article Snippet: RNA isolation and quantitative real-time PCR Excised wounds together with epidermal margins were mechanically dissociated in 500 μl trizol reagent using a power-driven homogenizer (OMNI International, Marietta, GA, USA), and total RNA (0.1–0.5 μg RNA/mg tissue) was extracted using the RNeasy lipid tissue Mini Kit (Qiagen). .. Quantitative differences in gene expression were determined by real-time quantitative PCR using SYBR-Green PCR master mix (Qiagen) and a spectrofluorometric thermal cycler (Mx3000P from Stratagene, La Jolla, CA, USA).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Dietary Salt Restriction Improves Cardiac and Adipose Tissue Pathology Independently of Obesity in a Rat Model of Metabolic Syndrome
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    DNA Extraction:

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    Article Title: A Viral Discovery Methodology for Clinical Biopsy Samples Utilising Massively Parallel Next Generation Sequencing
    Article Snippet: No Enrichment Total RNA extraction was performed on liver tissue (Liver biopsy or 2 mm3 ) using the RNeasy Lipid Tissue Mini Kit (Qiagen) according to the manufacturer's instructions with on column DNAse treatment. .. Total DNA extraction was performed on liver tissue (2 mm3 or from nuclear pellet) using the QIAamp DNA Mini Kit (Qiagen) and according to the manufacturer's instructions.

    Fluorescence:

    Article Title: Sustained Exendin-4 Secretion through Gene Therapy Targeting Salivary Glands in Two Different Rodent Models of Obesity/Type 2 Diabetes
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    Isolation:

    Article Title: Agouti Revisited: Transcript Quantification of the ASIP Gene in Bovine Tissues Related to Protein Expression and Localization
    Article Snippet: .. For RNA isolation, adipose tissues were homogenised in Qiazol lysis reagent (Qiagen, Hilden, Germany) with a Polytron homogenizer (Kinematica AG, Littau-Luzern, Switzerland) and total RNA was extracted with the RNeasy Lipid Tissue Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. .. Muscle tissue was homogenized using the Xiril Dispomix (Xiril, Hombrechtikon, Switzerland) and Qiazol Lysis Reagent (Qiagen, Hilden, Germany) as described by the manufacturer.

    Article Title: Dietary Salt Restriction Improves Cardiac and Adipose Tissue Pathology Independently of Obesity in a Rat Model of Metabolic Syndrome
    Article Snippet: Quantitative Reverse Transcription Polymerase Chain Reaction (RT‐PCR) Analysis Total RNA was extracted from LV tissue and treated with DNase with the use of a spin‐vacuum isolation kit (Promega, Madison, WI). .. Total RNA was extracted from adipose tissue homogenized with QIAzol reagent with the use of an RNeasy Lipid Tissue Mini Kit for adipose tissue (Qiagen, Hilden, Germany).

    Article Title: Indirect Effects of Glucagon-Like Peptide-1 Receptor Agonist Exendin-4 on the Peripheral Circadian Clocks in Mice
    Article Snippet: .. RNA extraction and real-time quantitative PCR Total RNA isolated from the peripheral tissues, using an RNeasy Mini Kit or an RNeasy Lipid Tissue Mini Kit (Qiagen, Valencia, CA), was reverse-transcribed using a PrimeScript RT reagent Kit (Takara Bio, Otsu, Japan). .. SCN samples were directly reverse-transcribed using a TaqMan Fast Cells-to-Ct Kit (Life Technologies, Carlsbad, CA).

    Article Title: mRNA expression of diacylglycerol kinase isoforms in insulin-sensitive tissues: effects of obesity and insulin resistance
    Article Snippet: .. Total RNA in adipose tissue and liver was isolated using RNeasy Lipid Tissue Mini Kit and RNeasy Mini Kit (Qiagen), respectively. .. A deoxyribonuclease I (QIAGEN) digestion step was included to eliminate DNA contamination.

    Article Title: BeeDoctor, a Versatile MLPA-Based Diagnostic Tool for Screening Bee Viruses
    Article Snippet: .. In order to have a better recovery of negative strand intermediates from replicating viruses, RNA was isolated with RNeasy Lipid Tissue Mini kit. .. No band of the correct size was obtained in the RT-free controls, for either the positive-strand or negative-strand MLPA reaction , showing that the NAD-dependent ligase-65 used in RT-MLPA cannot ligate DNA probe oligonucleotides that are hybridized to RNA.

    Purification:

    Article Title: Agouti Revisited: Transcript Quantification of the ASIP Gene in Bovine Tissues Related to Protein Expression and Localization
    Article Snippet: For RNA isolation, adipose tissues were homogenised in Qiazol lysis reagent (Qiagen, Hilden, Germany) with a Polytron homogenizer (Kinematica AG, Littau-Luzern, Switzerland) and total RNA was extracted with the RNeasy Lipid Tissue Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. .. The RNA was isolated and purified with NucleoSpin Extract II reagent (Macherey-Nagel, Dueren, Germany) according to manufacturer's guidelines.

    Article Title: Obesity and prostate cancer: gene expression signature of human periprostatic adipose tissue
    Article Snippet: .. RNA extraction, microarray hybridization and data processing Total RNA was extracted from PP adipose tissue samples after homogenization with an ULTRA-TURRAX T25 basic (IKA Werke GmbH, Staufen, Germany) in QIAzol reagent (Qiagen, Valencia, CA, USA) and purified through columns (RNeasy Lipid Tissue Mini kit, Qiagen) with DNase I treatment (RNase-free DNase set, Qiagen). .. Integrity and purity of RNA were assessed by on-chip electrophoresis using Experion (BioRad, Hercules, CA, USA).

    Sequencing:

    Article Title: Dietary Salt Restriction Improves Cardiac and Adipose Tissue Pathology Independently of Obesity in a Rat Model of Metabolic Syndrome
    Article Snippet: Total RNA was extracted from adipose tissue homogenized with QIAzol reagent with the use of an RNeasy Lipid Tissue Mini Kit for adipose tissue (Qiagen, Hilden, Germany). .. After RT, real‐time PCR analysis was performed with the use of a Prism 7000 Sequence Detector (Perkin‐Elmer, Wellesley, MA) and with primers and TaqMan probes specific for rat cDNAs encoding atrial natriuretic peptide, brain natriuretic peptide, β‐myosin heavy chain, collagen type I or type III, fibronectin, the p22phox , gp91phox , p47phox , p67phox , and Rac1 subunits of NADPH oxidase, monocyte chemoattractant protein (MCP)‐1, osteopontin, cyclooxygenase (COX)‐2, angiotensin‐converting enzyme, the angiotensin II type 1A receptor, the mineralocorticoid receptor (MR), serum‐ and glucocorticoid‐regulated kinase 1, TNF‐α, or IL‐6.

    Article Title: Selected imprinting of INS in the marsupial
    Article Snippet: RNA and DNA extraction and RT-PCR Individual genotypes were identified using PCR and direct sequencing of genomic DNA extracted from approximately 20 mg snap-frozen tissue using a Wizard Genomic DNA Purification Kit (Promega, Madison, WI, USA). .. Total RNA was extracted from mammary glands using the RNeasy Lipid Tissue Mini Kit (Qiagen,Hilden, Germany) or from other tissues using Tri-Reagent (Ambion, Austin, Texas, USA) as described by the manufacturer with a final elution of RNA in 60 to 80 μl RNAsecure H2 O (a dilution of 1/24 μl RNAsecure to water;Ambion).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Sustained Exendin-4 Secretion through Gene Therapy Targeting Salivary Glands in Two Different Rodent Models of Obesity/Type 2 Diabetes
    Article Snippet: RNA was extracted using RNeasy Lipid Tissue Mini Kit (QIAGEN GmbH, Hilden, Germany) according to the manufacturer’s instructions. .. Real Time quantitative PCR was carried out on DNA Engine Opticon™ 2 Continuous Fluorescence Detection System (MJ Research, MA, USA), using Platinum® SYBR® Green qPCR SuperMix-UDG (Invitrogen Corporation, CA, USA) and 300 nM specific primers for each gene : 18 s forward 5′-CGG CTA CCA CAT CCA AGG AA-3′, reverse 5′-GCT GGA ATT ACC GCG GCT-3′ ; leptin forward: 5′-TCC AGA AAG TCC AGG ATG ACA C-3′, reverse: 5′-CAC ATT TTG GGA AGG CAG G-3′; adiponectin forward: 5′-ACA ATG GCA CAC CAG GCC GTG A-3′, reverse: AGC GGC TTC TCC AGG CTC TCC TTT-3′.

    Mouse Assay:

    Article Title: Sustained Exendin-4 Secretion through Gene Therapy Targeting Salivary Glands in Two Different Rodent Models of Obesity/Type 2 Diabetes
    Article Snippet: Visceral Adipose Tissue Adipokines Profile: RNA Extraction and Real Time PCR Determinations Total RNA was extracted from 50 mg of mice visceral adipose tissue. .. RNA was extracted using RNeasy Lipid Tissue Mini Kit (QIAGEN GmbH, Hilden, Germany) according to the manufacturer’s instructions.

    Electrophoresis:

    Article Title: Obesity and prostate cancer: gene expression signature of human periprostatic adipose tissue
    Article Snippet: RNA extraction, microarray hybridization and data processing Total RNA was extracted from PP adipose tissue samples after homogenization with an ULTRA-TURRAX T25 basic (IKA Werke GmbH, Staufen, Germany) in QIAzol reagent (Qiagen, Valencia, CA, USA) and purified through columns (RNeasy Lipid Tissue Mini kit, Qiagen) with DNase I treatment (RNase-free DNase set, Qiagen). .. Integrity and purity of RNA were assessed by on-chip electrophoresis using Experion (BioRad, Hercules, CA, USA).

    RNA Extraction:

    Article Title: Dextran sulfate sodium-induced colitis alters stress-associated behaviour and neuropeptide gene expression in the amygdala-hippocampus network of mice
    Article Snippet: Real-time reverse transcription PCR Brain tissues used for real-time reverse transcription PCR were homogenized with a Precellys 24 homogenizer (Peqlab Biotechnology, Polling, Austria) and RNA was extracted with the RNeasy Lipid Tissue Mini kit (Qiagen, Vienna, Austria). .. The RNA extraction method was tested for quality on the BioAnalyzer BA2100 (Agilent, Foster City, CA, USA) with the RNA 6000 Nano LabChip Kit (catalogue number 5067-1511, Agilent).

    Article Title: Obesity and prostate cancer: gene expression signature of human periprostatic adipose tissue
    Article Snippet: .. RNA extraction, microarray hybridization and data processing Total RNA was extracted from PP adipose tissue samples after homogenization with an ULTRA-TURRAX T25 basic (IKA Werke GmbH, Staufen, Germany) in QIAzol reagent (Qiagen, Valencia, CA, USA) and purified through columns (RNeasy Lipid Tissue Mini kit, Qiagen) with DNase I treatment (RNase-free DNase set, Qiagen). .. Integrity and purity of RNA were assessed by on-chip electrophoresis using Experion (BioRad, Hercules, CA, USA).

    Article Title: Indirect Effects of Glucagon-Like Peptide-1 Receptor Agonist Exendin-4 on the Peripheral Circadian Clocks in Mice
    Article Snippet: .. RNA extraction and real-time quantitative PCR Total RNA isolated from the peripheral tissues, using an RNeasy Mini Kit or an RNeasy Lipid Tissue Mini Kit (Qiagen, Valencia, CA), was reverse-transcribed using a PrimeScript RT reagent Kit (Takara Bio, Otsu, Japan). .. SCN samples were directly reverse-transcribed using a TaqMan Fast Cells-to-Ct Kit (Life Technologies, Carlsbad, CA).

    Article Title: Sustained Exendin-4 Secretion through Gene Therapy Targeting Salivary Glands in Two Different Rodent Models of Obesity/Type 2 Diabetes
    Article Snippet: Paragraph title: Visceral Adipose Tissue Adipokines Profile: RNA Extraction and Real Time PCR Determinations ... RNA was extracted using RNeasy Lipid Tissue Mini Kit (QIAGEN GmbH, Hilden, Germany) according to the manufacturer’s instructions.

    Article Title: A Viral Discovery Methodology for Clinical Biopsy Samples Utilising Massively Parallel Next Generation Sequencing
    Article Snippet: .. No Enrichment Total RNA extraction was performed on liver tissue (Liver biopsy or 2 mm3 ) using the RNeasy Lipid Tissue Mini Kit (Qiagen) according to the manufacturer's instructions with on column DNAse treatment. .. Total DNA extraction was performed on liver tissue (2 mm3 or from nuclear pellet) using the QIAamp DNA Mini Kit (Qiagen) and according to the manufacturer's instructions.

    Agarose Gel Electrophoresis:

    Article Title: Selected imprinting of INS in the marsupial
    Article Snippet: Total RNA was extracted from mammary glands using the RNeasy Lipid Tissue Mini Kit (Qiagen,Hilden, Germany) or from other tissues using Tri-Reagent (Ambion, Austin, Texas, USA) as described by the manufacturer with a final elution of RNA in 60 to 80 μl RNAsecure H2 O (a dilution of 1/24 μl RNAsecure to water;Ambion). .. Total RNA was DNase treated (DNA-free™; Ambion) to remove contaminating DNA, run on a 1% agarose gel to assess the quality, quantified with a nano-spectrometer (NanoDrop ND-1000 Spectrophotometer; NanoDrop Technologies Inc., Wilmington, DE, USA) and cDNA was synthesised using the SuperScript III First Strand Synthesis System for RT-PCR (Invitrogen, Carisbad, CA, USA).

    Homogenization:

    Article Title: Obesity and prostate cancer: gene expression signature of human periprostatic adipose tissue
    Article Snippet: .. RNA extraction, microarray hybridization and data processing Total RNA was extracted from PP adipose tissue samples after homogenization with an ULTRA-TURRAX T25 basic (IKA Werke GmbH, Staufen, Germany) in QIAzol reagent (Qiagen, Valencia, CA, USA) and purified through columns (RNeasy Lipid Tissue Mini kit, Qiagen) with DNase I treatment (RNase-free DNase set, Qiagen). .. Integrity and purity of RNA were assessed by on-chip electrophoresis using Experion (BioRad, Hercules, CA, USA).

    Article Title: Sustained Exendin-4 Secretion through Gene Therapy Targeting Salivary Glands in Two Different Rodent Models of Obesity/Type 2 Diabetes
    Article Snippet: Briefly, tissue samples were collected, immediately snap frozen in liquid nitrogen and disrupted by homogenization in QIAzol Lysis Reagent using the TissueLyser (QIAGEN GmbH, Hilden, Germany). .. RNA was extracted using RNeasy Lipid Tissue Mini Kit (QIAGEN GmbH, Hilden, Germany) according to the manufacturer’s instructions.

    Spectrophotometry:

    Article Title: Agouti Revisited: Transcript Quantification of the ASIP Gene in Bovine Tissues Related to Protein Expression and Localization
    Article Snippet: For RNA isolation, adipose tissues were homogenised in Qiazol lysis reagent (Qiagen, Hilden, Germany) with a Polytron homogenizer (Kinematica AG, Littau-Luzern, Switzerland) and total RNA was extracted with the RNeasy Lipid Tissue Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. .. RNA was quantified with a NanoDrop ND-1000 spectrophotometer (Peqlab, Erlangen, Germany).

    Article Title: Perturbations of Fibroblast Growth Factors 19 and 21 in Type 2 Diabetes
    Article Snippet: Gene expression using real-time quantitative qPCR RNA preparations from liver biospecimens taken during RYGB surgery were performed using a kit (RNeasy Lipid Tissue Mini Kit, Qiagen, Valencia, CA) and as we have previously described [ ]. .. RNA quality evaluation and quantitation was performed using a Nanodrop ND-1000 spectrophotometer (Thermo Scientific, Wilmington, DE).

    Article Title: Regulation of energy metabolism during social interactions in rainbow trout: a role for AMP-activated protein kinase
    Article Snippet: Total RNA was extracted from liver or muscle tissue using an RNeasy Lipid Tissue Mini Kit (Qiagen, Toronto, ON, Canada) according to the manufacturer’s instructions. .. Extracted total RNA was quantified using a spectrophotometer (Nanodrop ND-2000c UV-Vis, Thermo Fisher Scientific). cDNA was synthesized using a QuantiTect reverse-transcription kit (Qiagen) according to the manufacturer’s protocol.

    Article Title: Selected imprinting of INS in the marsupial
    Article Snippet: Total RNA was extracted from mammary glands using the RNeasy Lipid Tissue Mini Kit (Qiagen,Hilden, Germany) or from other tissues using Tri-Reagent (Ambion, Austin, Texas, USA) as described by the manufacturer with a final elution of RNA in 60 to 80 μl RNAsecure H2 O (a dilution of 1/24 μl RNAsecure to water;Ambion). .. Total RNA was DNase treated (DNA-free™; Ambion) to remove contaminating DNA, run on a 1% agarose gel to assess the quality, quantified with a nano-spectrometer (NanoDrop ND-1000 Spectrophotometer; NanoDrop Technologies Inc., Wilmington, DE, USA) and cDNA was synthesised using the SuperScript III First Strand Synthesis System for RT-PCR (Invitrogen, Carisbad, CA, USA).

    Article Title: mRNA expression of diacylglycerol kinase isoforms in insulin-sensitive tissues: effects of obesity and insulin resistance
    Article Snippet: Total RNA in adipose tissue and liver was isolated using RNeasy Lipid Tissue Mini Kit and RNeasy Mini Kit (Qiagen), respectively. .. RNA concentration and purity were determined with a Nanodrop ND-1000 spectrophotometer (Nano-Drop Technologies, Wilmington, DE, USA) and all samples had a 260/280 and a 260/230 ratio > 2.0 indicating pure RNA.

    Concentration Assay:

    Article Title: Dextran sulfate sodium-induced colitis alters stress-associated behaviour and neuropeptide gene expression in the amygdala-hippocampus network of mice
    Article Snippet: Real-time reverse transcription PCR Brain tissues used for real-time reverse transcription PCR were homogenized with a Precellys 24 homogenizer (Peqlab Biotechnology, Polling, Austria) and RNA was extracted with the RNeasy Lipid Tissue Mini kit (Qiagen, Vienna, Austria). .. After extraction, the RNA concentration was measured and 1 μg RNA/sample submitted to a reverse transcription reaction using the high capacity cDNA reverse transcription kit (Applied Biosystems, Foster City, CA, USA).

    Article Title: mRNA expression of diacylglycerol kinase isoforms in insulin-sensitive tissues: effects of obesity and insulin resistance
    Article Snippet: Total RNA in adipose tissue and liver was isolated using RNeasy Lipid Tissue Mini Kit and RNeasy Mini Kit (Qiagen), respectively. .. RNA concentration and purity were determined with a Nanodrop ND-1000 spectrophotometer (Nano-Drop Technologies, Wilmington, DE, USA) and all samples had a 260/280 and a 260/230 ratio > 2.0 indicating pure RNA.

    DNA Purification:

    Article Title: Selected imprinting of INS in the marsupial
    Article Snippet: RNA and DNA extraction and RT-PCR Individual genotypes were identified using PCR and direct sequencing of genomic DNA extracted from approximately 20 mg snap-frozen tissue using a Wizard Genomic DNA Purification Kit (Promega, Madison, WI, USA). .. Total RNA was extracted from mammary glands using the RNeasy Lipid Tissue Mini Kit (Qiagen,Hilden, Germany) or from other tissues using Tri-Reagent (Ambion, Austin, Texas, USA) as described by the manufacturer with a final elution of RNA in 60 to 80 μl RNAsecure H2 O (a dilution of 1/24 μl RNAsecure to water;Ambion).

    Lysis:

    Article Title: Agouti Revisited: Transcript Quantification of the ASIP Gene in Bovine Tissues Related to Protein Expression and Localization
    Article Snippet: .. For RNA isolation, adipose tissues were homogenised in Qiazol lysis reagent (Qiagen, Hilden, Germany) with a Polytron homogenizer (Kinematica AG, Littau-Luzern, Switzerland) and total RNA was extracted with the RNeasy Lipid Tissue Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. .. Muscle tissue was homogenized using the Xiril Dispomix (Xiril, Hombrechtikon, Switzerland) and Qiazol Lysis Reagent (Qiagen, Hilden, Germany) as described by the manufacturer.

    Article Title: Sustained Exendin-4 Secretion through Gene Therapy Targeting Salivary Glands in Two Different Rodent Models of Obesity/Type 2 Diabetes
    Article Snippet: Briefly, tissue samples were collected, immediately snap frozen in liquid nitrogen and disrupted by homogenization in QIAzol Lysis Reagent using the TissueLyser (QIAGEN GmbH, Hilden, Germany). .. RNA was extracted using RNeasy Lipid Tissue Mini Kit (QIAGEN GmbH, Hilden, Germany) according to the manufacturer’s instructions.

    other:

    Article Title: P-Selectin Glycoprotein Ligand-1 Deficiency Is Protective Against Obesity-Related Insulin Resistance
    Article Snippet: Total RNA was extracted from each specimen of eWAT and liver using the RNeasy Lipid Tissue Mini Kit or RNeasy Plus Mini Kit following the instructions provided by the manufacturer (Qiagen, Valencia, CA).

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    Qiagen qrt pcr assay rneasy lipid tissue mini kit
    mRNA expression levels of Bcl-2 , Bax , and Cyt-c normalized to the transcription levels of β-actin and GAPDH . The <t>qRT-PCR</t> analysis was performed on COLO205 cells treated with 5.2 ± 0.8 μg/mL MGR. The experiment was done in triplicates, and data are expressed as mean ± SD. *Significant difference from control ( P
    Qrt Pcr Assay Rneasy Lipid Tissue Mini Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 95/100, based on 125 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mRNA expression levels of Bcl-2 , Bax , and Cyt-c normalized to the transcription levels of β-actin and GAPDH . The qRT-PCR analysis was performed on COLO205 cells treated with 5.2 ± 0.8 μg/mL MGR. The experiment was done in triplicates, and data are expressed as mean ± SD. *Significant difference from control ( P

    Journal: OncoTargets and therapy

    Article Title: Phytochemical analysis and antioxidant and anticancer activities of mastic gum resin from Pistacia atlantica subspecies kurdica

    doi: 10.2147/OTT.S170827

    Figure Lengend Snippet: mRNA expression levels of Bcl-2 , Bax , and Cyt-c normalized to the transcription levels of β-actin and GAPDH . The qRT-PCR analysis was performed on COLO205 cells treated with 5.2 ± 0.8 μg/mL MGR. The experiment was done in triplicates, and data are expressed as mean ± SD. *Significant difference from control ( P

    Article Snippet: qRT-PCR assay RNeasy® lipid tissue mini kit (Qiagen NV, Venlo, the Netherlands) was used to extract total RNA from COLO205 cells prior to conducting qRT-PCR analysis.

    Techniques: Expressing, Quantitative RT-PCR

    Phosphacan accumulation is impaired in the glial scar of Egr-1-deficient mice. A and B: Measurement of GFAP ( A ) and phosphacan ( B ) RNA expression levels 10 days after permanent occlusion of the MCA. RNA samples were isolated from wild-type (+/+) and Egr-1-deficient (−/−) mice and from the infarcted, left brain (L) and contralateral, right brain (R) hemispheres. Results were normalized to β-actin RNA expression levels setting arbitrarily the value of one wild-type sample from the right hemisphere as 100. Statistical analysis was performed with the Mann-Whitney rank sum test. Wild-type and Egr-1-deficient mice show comparable GFAP induction. Egr-1 −/− mice show a statistically significant reduction of phosphacan levels in the infarcted hemisphere compared with the contralateral site ( P = 0.001). The expression levels of phosphacan are also statistically significant different between the infarcted hemispheres of Egr-1 −/− mice and their wild-type littermates ( P ≤ 0.001). In contrast, there is no statistically significant change of phosphacan expression between both hemispheres ( P = 0.163) in wild-type mice and between noninfarcted hemispheres of WT and KO mice ( P = 0.380). C: Western blot analysis of brain tissue isolated from the glial scar 10 days after occlusion of the MCA and control (c) tissue from the contralateral brain hemisphere. Quantitative image analysis demonstrates that GFAP up-regulation after stroke is comparable in Egr-1 −/− and wild-type brain tissue, ie, 4.4 times in wild-type samples (SD, 0.6) and 4.7 times in Egr-1 −/− mice (SD, 1.8). The levels of the two phosphacan isoforms detected by the KAF13 antibody, namely the secreted form of phosphacan (Pcan) as well as the transmembrane isoform receptor protein tyrosine phosphatase β (RPTPβs) are up-regulated in the glial scar of wild types by 1.49-fold (SD, 0.14), but not in the glial scar of Egr-1-deficient animals (0.89-fold; SD, 0.26). β-Tubulin serves as loading control. Protein sizes are indicated in kDa.

    Journal: The American Journal of Pathology

    Article Title: Egr-1 Regulates Expression of the Glial Scar Component Phosphacan in Astrocytes after Experimental Stroke

    doi: 10.2353/ajpath.2008.070648

    Figure Lengend Snippet: Phosphacan accumulation is impaired in the glial scar of Egr-1-deficient mice. A and B: Measurement of GFAP ( A ) and phosphacan ( B ) RNA expression levels 10 days after permanent occlusion of the MCA. RNA samples were isolated from wild-type (+/+) and Egr-1-deficient (−/−) mice and from the infarcted, left brain (L) and contralateral, right brain (R) hemispheres. Results were normalized to β-actin RNA expression levels setting arbitrarily the value of one wild-type sample from the right hemisphere as 100. Statistical analysis was performed with the Mann-Whitney rank sum test. Wild-type and Egr-1-deficient mice show comparable GFAP induction. Egr-1 −/− mice show a statistically significant reduction of phosphacan levels in the infarcted hemisphere compared with the contralateral site ( P = 0.001). The expression levels of phosphacan are also statistically significant different between the infarcted hemispheres of Egr-1 −/− mice and their wild-type littermates ( P ≤ 0.001). In contrast, there is no statistically significant change of phosphacan expression between both hemispheres ( P = 0.163) in wild-type mice and between noninfarcted hemispheres of WT and KO mice ( P = 0.380). C: Western blot analysis of brain tissue isolated from the glial scar 10 days after occlusion of the MCA and control (c) tissue from the contralateral brain hemisphere. Quantitative image analysis demonstrates that GFAP up-regulation after stroke is comparable in Egr-1 −/− and wild-type brain tissue, ie, 4.4 times in wild-type samples (SD, 0.6) and 4.7 times in Egr-1 −/− mice (SD, 1.8). The levels of the two phosphacan isoforms detected by the KAF13 antibody, namely the secreted form of phosphacan (Pcan) as well as the transmembrane isoform receptor protein tyrosine phosphatase β (RPTPβs) are up-regulated in the glial scar of wild types by 1.49-fold (SD, 0.14), but not in the glial scar of Egr-1-deficient animals (0.89-fold; SD, 0.26). β-Tubulin serves as loading control. Protein sizes are indicated in kDa.

    Article Snippet: Total RNA was extracted and prepared from cultured cells as well as brain hemispheres (ipsilateral and contralateral) 10 days after permanent occlusion of the MCA using the RNA lipid tissue mini kit (Qiagen, Hilden, Germany).

    Techniques: Mouse Assay, RNA Expression, Isolation, MANN-WHITNEY, Expressing, Western Blot

    RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.

    Journal: PLoS ONE

    Article Title: Gene Expression in the Spinal Cord in Female Lewis Rats with Experimental Autoimmune Encephalomyelitis Induced with Myelin Basic Protein

    doi: 10.1371/journal.pone.0048555

    Figure Lengend Snippet: RT- PCR validation of 5 genes expressed differentially in the spinal cords of Lewis rats with MBP induced EAE. Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit. RNA quality analysis was carried out on the BioRadExperion automated electrophoresis system. All preparations used in both assays had RNA quality indicator (RQI) values of > 9.5. For RT-PCR, total RNA was reverse transcribed and amplified as described in the methods. Analysis of selected genes up or down regulated at the peak of disease in EAE. Bars represent the average fold change between expression in the spinal cord level at peak of disease compared to normal healthy animals (+/− SEMs, Microarray n = 4, RT-PCR n = 8). Dark columns represent fold change derived from the microarray data. Similar amplification patterns were obtained from RT-PCR amplification of the same total RNA samples and a second set of 4 animals samples at an identical time point.

    Article Snippet: Tissue samples were snap frozen in liquid nitrogen and stored at −80°C prior to total RNA preparation using the QIAGEN RNeasy Lipid tissue kit.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Electrophoresis, Amplification, Expressing, Microarray, Derivative Assay