rlt lysis buffer provided in the rneasy kit qiagen hilden germany  (Qiagen)


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    Name:
    Buffer RLT
    Description:
    For lysis of cells and tissues before RNA isolation. Kit contents: Qiagen Buffer RLT, 220mL, For RNeasy Lysis Buffer for Lysing Cells and Tissues Prior to RNA Isolation and Simultaneous RNA/DNA/Protein Isolation.
    Catalog Number:
    79216
    Price:
    None
    Category:
    Buffer RLT
    Score:
    85
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    Structured Review

    Qiagen rlt lysis buffer provided in the rneasy kit qiagen hilden germany
    Buffer RLT
    For lysis of cells and tissues before RNA isolation. Kit contents: Qiagen Buffer RLT, 220mL, For RNeasy Lysis Buffer for Lysing Cells and Tissues Prior to RNA Isolation and Simultaneous RNA/DNA/Protein Isolation.
    https://www.bioz.com/result/rlt lysis buffer provided in the rneasy kit qiagen hilden germany/product/Qiagen
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    rlt lysis buffer provided in the rneasy kit qiagen hilden germany - by Bioz Stars, 2019-10
    99/100 stars

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    Related Articles

    Methylation Sequencing:

    Article Title: Unique cell-type specific patterns of DNA methylation in the root meristem
    Article Snippet: Sorted cells were collected directly into specific lysis buffers that were compatible with downstream applications. .. Cells used for bisulfite sequencing, mRNA-seq, smRNA-seq were lysed in Buffer AP1 (Qiagen), Buffer RLT (Qiagen), Trizol (Invitrogen). .. All samples were immediately stored at −80 °C until gDNA and RNA was extracted using DNeasy Plant mini kit (Qiagen) and RNeasy Plant mini kit (Qiagen) or Trizol, respectively.

    Centrifugation:

    Article Title: Allele-specific transcription factor binding to common and rare variants associated with disease and gene expression
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were purified from healthy donor buffy coats (Department of Transfusion Medicine, Uppsala University Hospital, Sweden) using Ficoll density-gradient centrifugation. .. The harvested cells were stored in RLT buffer (QIAGEN) at −80 °C.

    Article Title: In vivo genome-wide profiling reveals a tissue-specific role for 5-formylcytosine
    Article Snippet: Tissues were collected from timed matings with the day of the copulation plug as E0.5. .. Embryonic tissues were lysed in RLT plus buffer (Qiagen), homogenised via centrifugation through a QIAshreddred column (Qiagen) at full speed for 2 min in a microcentrifuge. .. DNA and RNA were isolated using AllPrep DNA/RNA kit (Qiagen) following the manufacturer’s instructions.

    Amplification:

    Article Title: Targeted Mutation of NGN3 Gene Disrupts Pancreatic Endocrine Cell Development in Pigs
    Article Snippet: Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer. .. Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer.

    Cell Isolation:

    Article Title: Unique cell-type specific patterns of DNA methylation in the root meristem
    Article Snippet: Paragraph title: Cell isolation ... Cells used for bisulfite sequencing, mRNA-seq, smRNA-seq were lysed in Buffer AP1 (Qiagen), Buffer RLT (Qiagen), Trizol (Invitrogen).

    Quantitative RT-PCR:

    Article Title: Targeting Endothelial Adhesion Molecule Transcription for Treatment of Inflammatory Disease: A Proof-of-Concept Study
    Article Snippet: At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction. .. RNA concentration was determined by spectrophotometry on the NanoDrop 2000 instrument (ThermoFisher Scientific, Wilmington, DE).

    Article Title: Versatility of Biofilm Matrix Molecules in Staphylococcus epidermidis Clinical Isolates and Importance of Polysaccharide Intercellular Adhesin Expression during High Shear Stress
    Article Snippet: Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol. .. Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol.

    SYBR Green Assay:

    Article Title: Versatility of Biofilm Matrix Molecules in Staphylococcus epidermidis Clinical Isolates and Importance of Polysaccharide Intercellular Adhesin Expression during High Shear Stress
    Article Snippet: Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol. .. Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol.

    Incubation:

    Article Title: Genetic encoding of DNA nanostructures and their self-assembly in living bacteria
    Article Snippet: The RFP ratio is calculated by dividing the fluorescence with the expression of the RT by that in the absence of the RT (cells containing the same plasmid, including inducible system, but lacking the HIVRT genes). .. Cells were inoculated in 500 μl LB Miller Broth with antibiotics in a 96-well plate covered with a breathable membrane (AeraSeal, Excel Scientific) at 37 °C and 1,000 r.p.m. for 16 h. Overnight cultures were then diluted 1,000-fold by mixing 10 μl of the culture into 10 ml of LB Miller Broth containing the appropriate inducer and incubated at 37 °C and 250 r.p.m. for 18 h. After incubation, the DNA nanostructures were purified using the following protocol: (i) Cells were centrifuged at 5,000g for 7 min at 4 °C; (ii) the supernatant was removed; (iii) cells were resuspended in 200 μl of TE buffer (10 mM Tris-EDTA) containing 3 mg ml−1 of Lysozyme; (iv) 700 μl of RLT buffer (Qiagen, #79216) was added; (v) the resulting solution was centrifuged at 21,130 g for 2 min in order to remove the insoluble materials and the supernatant is transferred to a clean tube; (vi) 500 μl of 100% ethanol was added to the supernatant; (vii) 700 μl of the sample was transferred into a QIAquick Spin Column (Qiagen, #1018215) and centrifuged at 15,000g for 15 s; (viii) step vii was repeated until all the supernatant solution from step vi has passed through the same column tube (the flow through was discarded after each step); (ix) 700 μl of RW1 buffer (Qiagen, # 1053394) was added to the collection tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (x) 500 μl RPE buffer (Qiagen, # 1018013) was pipetted into the column tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (xi) 500 μl Buffer RPE (Mat. .. 1018013 Qiagen) was pipetted into the column tube and centrifuged for 2 min at 15,000g (the flow through was discarded); (xii) the empty column tube was centrifuged for 1 min at 15,000g ; (xiii) the column was placed into a clean tube and 50 μl of water was added; (xiv) the resulting solution was then incubated with RNase A (100 μg ml−1 , Qiagen) in the presence of 150 mM NaCl to recover the DNA–RNA chimera or without salt to recover just the ssDNA.

    Article Title: Allele-specific transcription factor binding to common and rare variants associated with disease and gene expression
    Article Snippet: The cells were stimulated with a phosphorothioate-modified CpG A oligonucleotide ODN2216 (CyberGene) at the concentration of 3 μg/ml and incubated for 5 h at 37 °C with 5 % CO2 . .. The harvested cells were stored in RLT buffer (QIAGEN) at −80 °C.

    Expressing:

    Article Title: A phylogenomic study quantifies competing mechanisms for pseudogenization in prokaryotes—The Mycobacterium leprae case
    Article Snippet: Paragraph title: Expression analysis ... Briefly, frozen tissue sections of nine lepromatous leprosy skin lesions, taken at the time of diagnosis after written consent was obtained and before any treatment was started, were lysed in Qiagen RLT buffer (Qiagen, Hilden, Germany) and homogenized with silica beads.

    Article Title: Targeted Mutation of NGN3 Gene Disrupts Pancreatic Endocrine Cell Development in Pigs
    Article Snippet: Paragraph title: Gene expression analysis using RT-PCR ... Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer.

    Article Title: Gene expression of INPP5F as an independent prognostic marker in fludarabine-based therapy of chronic lymphocytic leukemia
    Article Snippet: Paragraph title: Gene expression profiling ... CLL samples were positively enriched by magnetic cell sorting using CD19 microbeads and MACS columns (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and resuspended in RLT buffer (Qiagen, Crawley, West Sussex, UK).

    Article Title: S100A8 promotes migration and infiltration of inflammatory cells in acute anterior uveitis
    Article Snippet: Next, the iris-ciliary body tissue was isolated from the stabilized eye, and then homogenized with a rotor- stator homogenizer in buffer RLT (Qiagen). .. Next, the iris-ciliary body tissue was isolated from the stabilized eye, and then homogenized with a rotor- stator homogenizer in buffer RLT (Qiagen).

    Article Title: Does Caspase-6 Have a Role in Perinatal Brain Injury?
    Article Snippet: The entire supernatant was collected (1 ml) at 6, 24 or 48 h; 300 μl was used immediately for a lactate dehydrogenase assay (Roche, USA) and the remaining supernatant stored at −80°C until further analysis. .. Cells were washed with RNAse-free PBS and then lysed with 350-μl buffer RLT (Qiagen; containing β-mercaptoethanol at a concentration of 1:100), harvested and stored at −80°C for gene expression analysis. .. All time points were run in each independent experiment and all group experiments were performed a minimum of 3-6 independent times.

    Article Title: Versatility of Biofilm Matrix Molecules in Staphylococcus epidermidis Clinical Isolates and Importance of Polysaccharide Intercellular Adhesin Expression during High Shear Stress
    Article Snippet: Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol. .. Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol.

    Article Title: Cardiac Non-myocyte Cells Show Enhanced Pharmacological Function Suggestive of Contractile Maturity in Stem Cell Derived Cardiomyocyte Microtissues
    Article Snippet: Paragraph title: Gene expression analysis ... All primary cells were detached with prewarmed Accutase (Sigma, A6964) for 5 min at 37°C, 5% CO2 , centrifuged for 3 min at 1200 ×g before lysis in RLT buffer (Qiagen) and stored at −80 °C until processing.

    Real-time Polymerase Chain Reaction:

    Article Title: S100A8 promotes migration and infiltration of inflammatory cells in acute anterior uveitis
    Article Snippet: Paragraph title: Real-time PCR ... Next, the iris-ciliary body tissue was isolated from the stabilized eye, and then homogenized with a rotor- stator homogenizer in buffer RLT (Qiagen).

    Article Title: Targeting Endothelial Adhesion Molecule Transcription for Treatment of Inflammatory Disease: A Proof-of-Concept Study
    Article Snippet: At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction. .. At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction.

    Article Title: PEGylated IL-10 Activates Kupffer Cells to Control Hypercholesterolemia
    Article Snippet: Paragraph title: qPCR analysis ... Mouse livers were ground up in Buffer RLT (QIAGEN) with 10 μL β-mercaptoethanol (Sigma-Aldrich) using pestles (VWR), after which RNA was extracted using an RNeasy kit (QIAGEN) according to the manufacturer’s instructions.

    Article Title: Versatility of Biofilm Matrix Molecules in Staphylococcus epidermidis Clinical Isolates and Importance of Polysaccharide Intercellular Adhesin Expression during High Shear Stress
    Article Snippet: Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol. .. Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol.

    Flow Cytometry:

    Article Title: Genetic encoding of DNA nanostructures and their self-assembly in living bacteria
    Article Snippet: The RFP ratio is calculated by dividing the fluorescence with the expression of the RT by that in the absence of the RT (cells containing the same plasmid, including inducible system, but lacking the HIVRT genes). .. Cells were inoculated in 500 μl LB Miller Broth with antibiotics in a 96-well plate covered with a breathable membrane (AeraSeal, Excel Scientific) at 37 °C and 1,000 r.p.m. for 16 h. Overnight cultures were then diluted 1,000-fold by mixing 10 μl of the culture into 10 ml of LB Miller Broth containing the appropriate inducer and incubated at 37 °C and 250 r.p.m. for 18 h. After incubation, the DNA nanostructures were purified using the following protocol: (i) Cells were centrifuged at 5,000g for 7 min at 4 °C; (ii) the supernatant was removed; (iii) cells were resuspended in 200 μl of TE buffer (10 mM Tris-EDTA) containing 3 mg ml−1 of Lysozyme; (iv) 700 μl of RLT buffer (Qiagen, #79216) was added; (v) the resulting solution was centrifuged at 21,130 g for 2 min in order to remove the insoluble materials and the supernatant is transferred to a clean tube; (vi) 500 μl of 100% ethanol was added to the supernatant; (vii) 700 μl of the sample was transferred into a QIAquick Spin Column (Qiagen, #1018215) and centrifuged at 15,000g for 15 s; (viii) step vii was repeated until all the supernatant solution from step vi has passed through the same column tube (the flow through was discarded after each step); (ix) 700 μl of RW1 buffer (Qiagen, # 1053394) was added to the collection tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (x) 500 μl RPE buffer (Qiagen, # 1018013) was pipetted into the column tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (xi) 500 μl Buffer RPE (Mat. .. 1018013 Qiagen) was pipetted into the column tube and centrifuged for 2 min at 15,000g (the flow through was discarded); (xii) the empty column tube was centrifuged for 1 min at 15,000g ; (xiii) the column was placed into a clean tube and 50 μl of water was added; (xiv) the resulting solution was then incubated with RNase A (100 μg ml−1 , Qiagen) in the presence of 150 mM NaCl to recover the DNA–RNA chimera or without salt to recover just the ssDNA.

    Article Title: Versatility of Biofilm Matrix Molecules in Staphylococcus epidermidis Clinical Isolates and Importance of Polysaccharide Intercellular Adhesin Expression during High Shear Stress
    Article Snippet: Biofilms were grown at 37°C in 100% TSB under conditions of constant flow for 24 h at a rate of either 0.25 ml/min (“slow”) or 2 ml/min (“fast”). .. Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol. .. Cells were disrupted using a FastPrep FP120 device (Thermo Scientific).

    Cell Culture:

    Article Title: The transcription factors SOX9 and SOX5/SOX6 cooperate genome-wide through super-enhancers to drive chondrogenesis
    Article Snippet: RCS cells were harvested at 95% confluence in Buffer RLT (Qiagen). .. Total RNA was extracted and purified using RNeasy Mini Kit (Qiagen).

    Article Title: Allele-specific transcription factor binding to common and rare variants associated with disease and gene expression
    Article Snippet: The cells were cultured in 1 ml volumes in 24-well plates (Nunc) in macrophage serum-free medium (Life Technologies) at the concentration of 3 × 106 B cells/ml. .. The harvested cells were stored in RLT buffer (QIAGEN) at −80 °C.

    Article Title: Cardiac Non-myocyte Cells Show Enhanced Pharmacological Function Suggestive of Contractile Maturity in Stem Cell Derived Cardiomyocyte Microtissues
    Article Snippet: Adult left ventricle heart total RNA was obtained from one 21-year old normal male donor with no reported concomitant disease. hESC-CM total RNA was obtained from hESC-CMs cultured as described in . .. All primary cells were detached with prewarmed Accutase (Sigma, A6964) for 5 min at 37°C, 5% CO2 , centrifuged for 3 min at 1200 ×g before lysis in RLT buffer (Qiagen) and stored at −80 °C until processing.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Targeted Mutation of NGN3 Gene Disrupts Pancreatic Endocrine Cell Development in Pigs
    Article Snippet: Paragraph title: Gene expression analysis using RT-PCR ... Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer.

    Lactate Dehydrogenase Assay:

    Article Title: Does Caspase-6 Have a Role in Perinatal Brain Injury?
    Article Snippet: Cells were washed with RNAse-free PBS and then lysed with 350-μl buffer RLT (Qiagen; containing β-mercaptoethanol at a concentration of 1:100), harvested and stored at −80°C for gene expression analysis. .. Cells were washed with RNAse-free PBS and then lysed with 350-μl buffer RLT (Qiagen; containing β-mercaptoethanol at a concentration of 1:100), harvested and stored at −80°C for gene expression analysis.

    Generated:

    Article Title: Versatility of Biofilm Matrix Molecules in Staphylococcus epidermidis Clinical Isolates and Importance of Polysaccharide Intercellular Adhesin Expression during High Shear Stress
    Article Snippet: Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol. .. Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol.

    other:

    Article Title: Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies
    Article Snippet: The decrease of lysate volume (below 4 μl) during removal of cutout membranes from the tube can be adjusted with buffer RLT.

    Polymerase Chain Reaction:

    Article Title: Targeted Mutation of NGN3 Gene Disrupts Pancreatic Endocrine Cell Development in Pigs
    Article Snippet: Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer. .. Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer.

    Article Title: Targeting Endothelial Adhesion Molecule Transcription for Treatment of Inflammatory Disease: A Proof-of-Concept Study
    Article Snippet: Paragraph title: 2.3. RNA Isolation, Reverse Transcription, and Polymerase Chain Reaction ... At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction.

    Magnetic Cell Separation:

    Article Title: Gene expression of INPP5F as an independent prognostic marker in fludarabine-based therapy of chronic lymphocytic leukemia
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were obtained by Ficoll density gradient centrifugation. .. CLL samples were positively enriched by magnetic cell sorting using CD19 microbeads and MACS columns (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and resuspended in RLT buffer (Qiagen, Crawley, West Sussex, UK). .. In addition, as a validation set, PBMC samples (n =92) were processed into RLT buffer without CD19 enrichment.

    Nucleic Acid Electrophoresis:

    Article Title: Normalization with Corresponding Naïve Tissue Minimizes Bias Caused by Commercial Reverse Transcription Kits on Quantitative Real-Time PCR Results
    Article Snippet: For RNA extraction, brain tissue samples were lysed with RLT-buffer (Qiagen, Hilden, Germany) and homogenized with a MM300 mill mixer (Retsch, Haan, Germany) at 30 Hz for 2 minutes. .. Then, we isolated the total RNA from the lysed and homogenized cells by using a RNeasy Lipid Tissue Mini Kit (Qiagen) according to the manufacturer's instructions and eluted these with 30 μL to 50 μL RNase-free water.

    RNA Sequencing Assay:

    Article Title: A phylogenomic study quantifies competing mechanisms for pseudogenization in prokaryotes—The Mycobacterium leprae case
    Article Snippet: RNA sequencing data of leprosy skin lesions was analyzed from [ ]. .. Briefly, frozen tissue sections of nine lepromatous leprosy skin lesions, taken at the time of diagnosis after written consent was obtained and before any treatment was started, were lysed in Qiagen RLT buffer (Qiagen, Hilden, Germany) and homogenized with silica beads.

    Article Title: The transcription factors SOX9 and SOX5/SOX6 cooperate genome-wide through super-enhancers to drive chondrogenesis
    Article Snippet: Paragraph title: RNA-seq assay and data analysis ... RCS cells were harvested at 95% confluence in Buffer RLT (Qiagen).

    Isolation:

    Article Title: Targeted Mutation of NGN3 Gene Disrupts Pancreatic Endocrine Cell Development in Pigs
    Article Snippet: Pancreas tissue was isolated from fetal pigs at day 60 d.p.c. and immediately placed into RNALater Solution (Ambion). .. Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer.

    Article Title: Allele-specific transcription factor binding to common and rare variants associated with disease and gene expression
    Article Snippet: The B cells were isolated from PBMCs by positive selection using CD19+ B-cell isolation kit (Miltenyi Biotec) according to manufacturer’s instructions. .. The harvested cells were stored in RLT buffer (QIAGEN) at −80 °C.

    Article Title: Gene expression of INPP5F as an independent prognostic marker in fludarabine-based therapy of chronic lymphocytic leukemia
    Article Snippet: CLL samples were positively enriched by magnetic cell sorting using CD19 microbeads and MACS columns (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and resuspended in RLT buffer (Qiagen, Crawley, West Sussex, UK). .. CLL samples were positively enriched by magnetic cell sorting using CD19 microbeads and MACS columns (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and resuspended in RLT buffer (Qiagen, Crawley, West Sussex, UK).

    Article Title: A virosomal respiratory syncytial virus vaccine adjuvanted with monophosphoryl lipid A provides protection against viral challenge without priming for enhanced disease in cotton rats) A virosomal respiratory syncytial virus vaccine adjuvanted with monophosphoryl lipid A provides protection against viral challenge without priming for enhanced disease in cotton rats
    Article Snippet: Paragraph title: Lung RNA isolation ... Lung samples stored in RNA later (Qiagen) were transferred to tubes containing 300 μl RLT buffer (Qiagen) and homogenized using disposable pestles with a pestle motor (VWR, Amsterdam, the Netherlands).

    Article Title: S100A8 promotes migration and infiltration of inflammatory cells in acute anterior uveitis
    Article Snippet: The eye globe was enucleated and immediately submerged in RNA stabilization reagent (Qiagen, Hilden, Germany). .. Next, the iris-ciliary body tissue was isolated from the stabilized eye, and then homogenized with a rotor- stator homogenizer in buffer RLT (Qiagen). .. Total RNA was extracted using an RNeasy Protect Mini Kit (Qiagen) and treated with RNase- free DNase Set (Qiagen) to remove any residual genomic DNA. cDNA from each sample was obtained by reverse transcription with random hexamers using MultiScribe reverse transcriptase (Thermo Fisher Scientific, Waltham, MA).

    Article Title: Normalization with Corresponding Naïve Tissue Minimizes Bias Caused by Commercial Reverse Transcription Kits on Quantitative Real-Time PCR Results
    Article Snippet: Paragraph title: RNA isolation, quality control and cDNA synthesis ... For RNA extraction, brain tissue samples were lysed with RLT-buffer (Qiagen, Hilden, Germany) and homogenized with a MM300 mill mixer (Retsch, Haan, Germany) at 30 Hz for 2 minutes.

    Article Title: Targeting Endothelial Adhesion Molecule Transcription for Treatment of Inflammatory Disease: A Proof-of-Concept Study
    Article Snippet: Paragraph title: 2.3. RNA Isolation, Reverse Transcription, and Polymerase Chain Reaction ... At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction.

    Article Title: Versatility of Biofilm Matrix Molecules in Staphylococcus epidermidis Clinical Isolates and Importance of Polysaccharide Intercellular Adhesin Expression during High Shear Stress
    Article Snippet: Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol. .. Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol.

    Article Title: Suppression of ischemia in arterial occlusive disease by JNK-promoted native collateral artery development
    Article Snippet: Total RNA was extracted with TRIzol (Life Technologies) and was purified using an RNeasy kit (Qiagen). .. RNA from cells and other tissues homogenized in RLT buffer (Qiagen) was isolated using the RNeasy kit. .. We used purified RNA to prepare cDNA using The High Capacity Reverse Transcription Kit (Life Technologies).

    Article Title: Pathogenic Differences between Nipah Virus Bangladesh and Malaysia Strains in Primates: Implications for Antibody Therapy
    Article Snippet: Paragraph title: RNA Isolation ... RNAlater was completely removed, and tissues were homogenized in 600 μl RLT buffer (Qiagen) in a 2-mL cryovial using a tissue lyser (Qiagen) and ceramic beads.

    RNA Extraction:

    Article Title: Gene expression of INPP5F as an independent prognostic marker in fludarabine-based therapy of chronic lymphocytic leukemia
    Article Snippet: CLL samples were positively enriched by magnetic cell sorting using CD19 microbeads and MACS columns (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and resuspended in RLT buffer (Qiagen, Crawley, West Sussex, UK). .. CLL samples were positively enriched by magnetic cell sorting using CD19 microbeads and MACS columns (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and resuspended in RLT buffer (Qiagen, Crawley, West Sussex, UK).

    Article Title: Normalization with Corresponding Naïve Tissue Minimizes Bias Caused by Commercial Reverse Transcription Kits on Quantitative Real-Time PCR Results
    Article Snippet: The right upper regions were collected, frozen in liquid nitrogen and stored at -80°C. .. For RNA extraction, brain tissue samples were lysed with RLT-buffer (Qiagen, Hilden, Germany) and homogenized with a MM300 mill mixer (Retsch, Haan, Germany) at 30 Hz for 2 minutes. .. Then, we isolated the total RNA from the lysed and homogenized cells by using a RNeasy Lipid Tissue Mini Kit (Qiagen) according to the manufacturer's instructions and eluted these with 30 μL to 50 μL RNase-free water.

    Article Title: In vivo genome-wide profiling reveals a tissue-specific role for 5-formylcytosine
    Article Snippet: Paragraph title: DNA and RNA extraction ... Embryonic tissues were lysed in RLT plus buffer (Qiagen), homogenised via centrifugation through a QIAshreddred column (Qiagen) at full speed for 2 min in a microcentrifuge.

    Article Title: Targeting Endothelial Adhesion Molecule Transcription for Treatment of Inflammatory Disease: A Proof-of-Concept Study
    Article Snippet: The THP-1 human leukocyte line (American Type Culture Collection, Manassas, VA) was maintained in suspension in RPMI-1640 medium (ThermoFisher-GIBCO, Grand Island, NY) supplemented with 10% FBS and 0.05 mM 2-mercaptoethanol. .. At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction. .. Total RNA was extracted using the RNeasy mini kit (Qiagen), according to the manufacturer's instructions and including the optional on-column DNase treatment.

    Article Title: Pathogenic Differences between Nipah Virus Bangladesh and Malaysia Strains in Primates: Implications for Antibody Therapy
    Article Snippet: Immediately following sampling, 100 μl of blood was added to 600 μl of AVL viral lysis buffer (Qiagen) for RNA extraction. .. RNAlater was completely removed, and tissues were homogenized in 600 μl RLT buffer (Qiagen) in a 2-mL cryovial using a tissue lyser (Qiagen) and ceramic beads.

    Purification:

    Article Title: The transcription factors SOX9 and SOX5/SOX6 cooperate genome-wide through super-enhancers to drive chondrogenesis
    Article Snippet: RCS cells were harvested at 95% confluence in Buffer RLT (Qiagen). .. RCS cells were harvested at 95% confluence in Buffer RLT (Qiagen).

    Article Title: Genetic encoding of DNA nanostructures and their self-assembly in living bacteria
    Article Snippet: The RFP ratio is calculated by dividing the fluorescence with the expression of the RT by that in the absence of the RT (cells containing the same plasmid, including inducible system, but lacking the HIVRT genes). .. Cells were inoculated in 500 μl LB Miller Broth with antibiotics in a 96-well plate covered with a breathable membrane (AeraSeal, Excel Scientific) at 37 °C and 1,000 r.p.m. for 16 h. Overnight cultures were then diluted 1,000-fold by mixing 10 μl of the culture into 10 ml of LB Miller Broth containing the appropriate inducer and incubated at 37 °C and 250 r.p.m. for 18 h. After incubation, the DNA nanostructures were purified using the following protocol: (i) Cells were centrifuged at 5,000g for 7 min at 4 °C; (ii) the supernatant was removed; (iii) cells were resuspended in 200 μl of TE buffer (10 mM Tris-EDTA) containing 3 mg ml−1 of Lysozyme; (iv) 700 μl of RLT buffer (Qiagen, #79216) was added; (v) the resulting solution was centrifuged at 21,130 g for 2 min in order to remove the insoluble materials and the supernatant is transferred to a clean tube; (vi) 500 μl of 100% ethanol was added to the supernatant; (vii) 700 μl of the sample was transferred into a QIAquick Spin Column (Qiagen, #1018215) and centrifuged at 15,000g for 15 s; (viii) step vii was repeated until all the supernatant solution from step vi has passed through the same column tube (the flow through was discarded after each step); (ix) 700 μl of RW1 buffer (Qiagen, # 1053394) was added to the collection tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (x) 500 μl RPE buffer (Qiagen, # 1018013) was pipetted into the column tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (xi) 500 μl Buffer RPE (Mat. .. 1018013 Qiagen) was pipetted into the column tube and centrifuged for 2 min at 15,000g (the flow through was discarded); (xii) the empty column tube was centrifuged for 1 min at 15,000g ; (xiii) the column was placed into a clean tube and 50 μl of water was added; (xiv) the resulting solution was then incubated with RNase A (100 μg ml−1 , Qiagen) in the presence of 150 mM NaCl to recover the DNA–RNA chimera or without salt to recover just the ssDNA.

    Article Title: Allele-specific transcription factor binding to common and rare variants associated with disease and gene expression
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were purified from healthy donor buffy coats (Department of Transfusion Medicine, Uppsala University Hospital, Sweden) using Ficoll density-gradient centrifugation. .. The harvested cells were stored in RLT buffer (QIAGEN) at −80 °C.

    Article Title: Versatility of Biofilm Matrix Molecules in Staphylococcus epidermidis Clinical Isolates and Importance of Polysaccharide Intercellular Adhesin Expression during High Shear Stress
    Article Snippet: Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol. .. Biofilms were harvested by removing TSB from the flow cell chamber and resuspending biofilm cells in 900 µl of RLT buffer (Qiagen)–1% β-mercaptoethanol.

    Article Title: Suppression of ischemia in arterial occlusive disease by JNK-promoted native collateral artery development
    Article Snippet: Total RNA was extracted with TRIzol (Life Technologies) and was purified using an RNeasy kit (Qiagen). .. RNA from cells and other tissues homogenized in RLT buffer (Qiagen) was isolated using the RNeasy kit.

    Sequencing:

    Article Title: PEGylated IL-10 Activates Kupffer Cells to Control Hypercholesterolemia
    Article Snippet: Mouse livers were ground up in Buffer RLT (QIAGEN) with 10 μL β-mercaptoethanol (Sigma-Aldrich) using pestles (VWR), after which RNA was extracted using an RNeasy kit (QIAGEN) according to the manufacturer’s instructions. .. The purified RNA was used as template for RT-PCR using an RT2 First Strand kit (QIAGEN).

    FACS:

    Article Title: Gene expression of INPP5F as an independent prognostic marker in fludarabine-based therapy of chronic lymphocytic leukemia
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were obtained by Ficoll density gradient centrifugation. .. CLL samples were positively enriched by magnetic cell sorting using CD19 microbeads and MACS columns (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and resuspended in RLT buffer (Qiagen, Crawley, West Sussex, UK). .. In addition, as a validation set, PBMC samples (n =92) were processed into RLT buffer without CD19 enrichment.

    Article Title: Unique cell-type specific patterns of DNA methylation in the root meristem
    Article Snippet: Fluorescent Activated Cell Sorting (FACS) was performed using cell specific GFP lines as described previously . .. Cells used for bisulfite sequencing, mRNA-seq, smRNA-seq were lysed in Buffer AP1 (Qiagen), Buffer RLT (Qiagen), Trizol (Invitrogen).

    Blocking Assay:

    Article Title: Suppression of ischemia in arterial occlusive disease by JNK-promoted native collateral artery development
    Article Snippet: Hindlimb adductor and calf skeletal muscles were harvested en block, snap frozen in liquid nitrogen, and then pulverized into a powder using a CryoPREP impactor (Covaris (Woburn, MA)). .. RNA from cells and other tissues homogenized in RLT buffer (Qiagen) was isolated using the RNeasy kit.

    Lysis:

    Article Title: S100A8 promotes migration and infiltration of inflammatory cells in acute anterior uveitis
    Article Snippet: Leukocytes were isolated using SV RNA red blood cell lysis solution (Promega, Madison, WI), and total RNA was extracted using the SV total RNA isolation system (Promega). .. Next, the iris-ciliary body tissue was isolated from the stabilized eye, and then homogenized with a rotor- stator homogenizer in buffer RLT (Qiagen).

    Article Title: Unique cell-type specific patterns of DNA methylation in the root meristem
    Article Snippet: Cells used for bisulfite sequencing, mRNA-seq, smRNA-seq were lysed in Buffer AP1 (Qiagen), Buffer RLT (Qiagen), Trizol (Invitrogen). .. Cells used for bisulfite sequencing, mRNA-seq, smRNA-seq were lysed in Buffer AP1 (Qiagen), Buffer RLT (Qiagen), Trizol (Invitrogen).

    Article Title: Cardiac Non-myocyte Cells Show Enhanced Pharmacological Function Suggestive of Contractile Maturity in Stem Cell Derived Cardiomyocyte Microtissues
    Article Snippet: RNA was harvested following 72 h in culture as per the recommended manufacturer instructions. .. All primary cells were detached with prewarmed Accutase (Sigma, A6964) for 5 min at 37°C, 5% CO2 , centrifuged for 3 min at 1200 ×g before lysis in RLT buffer (Qiagen) and stored at −80 °C until processing. .. Microtissues were pooled and transferred into a falcon tube, centrifuged at 1200 ×g for 2 mins and re-suspended in prewarmed PBS.

    Article Title: Pathogenic Differences between Nipah Virus Bangladesh and Malaysia Strains in Primates: Implications for Antibody Therapy
    Article Snippet: Immediately following sampling, 100 μl of blood was added to 600 μl of AVL viral lysis buffer (Qiagen) for RNA extraction. .. RNAlater was completely removed, and tissues were homogenized in 600 μl RLT buffer (Qiagen) in a 2-mL cryovial using a tissue lyser (Qiagen) and ceramic beads.

    Agarose Gel Electrophoresis:

    Article Title: Targeted Mutation of NGN3 Gene Disrupts Pancreatic Endocrine Cell Development in Pigs
    Article Snippet: Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer. .. Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer.

    Mouse Assay:

    Article Title: Suppression of ischemia in arterial occlusive disease by JNK-promoted native collateral artery development
    Article Snippet: To isolate RNA from tissues, mice were perfusion cleared with PBS via the left ventricle. .. RNA from cells and other tissues homogenized in RLT buffer (Qiagen) was isolated using the RNeasy kit.

    Electrophoresis:

    Article Title: Targeted Mutation of NGN3 Gene Disrupts Pancreatic Endocrine Cell Development in Pigs
    Article Snippet: Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer. .. Approximately 30 mg of tissue was lysed using Qiagen buffer RLT with Betamercaptoethanol (BME) and polytron benchtop tissue homogenizer.

    Article Title: Genetic encoding of DNA nanostructures and their self-assembly in living bacteria
    Article Snippet: Cells were inoculated in 500 μl LB Miller Broth with antibiotics in a 96-well plate covered with a breathable membrane (AeraSeal, Excel Scientific) at 37 °C and 1,000 r.p.m. for 16 h. Overnight cultures were then diluted 1,000-fold by mixing 10 μl of the culture into 10 ml of LB Miller Broth containing the appropriate inducer and incubated at 37 °C and 250 r.p.m. for 18 h. After incubation, the DNA nanostructures were purified using the following protocol: (i) Cells were centrifuged at 5,000g for 7 min at 4 °C; (ii) the supernatant was removed; (iii) cells were resuspended in 200 μl of TE buffer (10 mM Tris-EDTA) containing 3 mg ml−1 of Lysozyme; (iv) 700 μl of RLT buffer (Qiagen, #79216) was added; (v) the resulting solution was centrifuged at 21,130 g for 2 min in order to remove the insoluble materials and the supernatant is transferred to a clean tube; (vi) 500 μl of 100% ethanol was added to the supernatant; (vii) 700 μl of the sample was transferred into a QIAquick Spin Column (Qiagen, #1018215) and centrifuged at 15,000g for 15 s; (viii) step vii was repeated until all the supernatant solution from step vi has passed through the same column tube (the flow through was discarded after each step); (ix) 700 μl of RW1 buffer (Qiagen, # 1053394) was added to the collection tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (x) 500 μl RPE buffer (Qiagen, # 1018013) was pipetted into the column tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (xi) 500 μl Buffer RPE (Mat. .. Cells were inoculated in 500 μl LB Miller Broth with antibiotics in a 96-well plate covered with a breathable membrane (AeraSeal, Excel Scientific) at 37 °C and 1,000 r.p.m. for 16 h. Overnight cultures were then diluted 1,000-fold by mixing 10 μl of the culture into 10 ml of LB Miller Broth containing the appropriate inducer and incubated at 37 °C and 250 r.p.m. for 18 h. After incubation, the DNA nanostructures were purified using the following protocol: (i) Cells were centrifuged at 5,000g for 7 min at 4 °C; (ii) the supernatant was removed; (iii) cells were resuspended in 200 μl of TE buffer (10 mM Tris-EDTA) containing 3 mg ml−1 of Lysozyme; (iv) 700 μl of RLT buffer (Qiagen, #79216) was added; (v) the resulting solution was centrifuged at 21,130 g for 2 min in order to remove the insoluble materials and the supernatant is transferred to a clean tube; (vi) 500 μl of 100% ethanol was added to the supernatant; (vii) 700 μl of the sample was transferred into a QIAquick Spin Column (Qiagen, #1018215) and centrifuged at 15,000g for 15 s; (viii) step vii was repeated until all the supernatant solution from step vi has passed through the same column tube (the flow through was discarded after each step); (ix) 700 μl of RW1 buffer (Qiagen, # 1053394) was added to the collection tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (x) 500 μl RPE buffer (Qiagen, # 1018013) was pipetted into the column tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (xi) 500 μl Buffer RPE (Mat.

    Article Title: Normalization with Corresponding Naïve Tissue Minimizes Bias Caused by Commercial Reverse Transcription Kits on Quantitative Real-Time PCR Results
    Article Snippet: For RNA extraction, brain tissue samples were lysed with RLT-buffer (Qiagen, Hilden, Germany) and homogenized with a MM300 mill mixer (Retsch, Haan, Germany) at 30 Hz for 2 minutes. .. Then, we isolated the total RNA from the lysed and homogenized cells by using a RNeasy Lipid Tissue Mini Kit (Qiagen) according to the manufacturer's instructions and eluted these with 30 μL to 50 μL RNase-free water.

    Negative Control:

    Article Title: Does Caspase-6 Have a Role in Perinatal Brain Injury?
    Article Snippet: Cells were washed with RNAse-free PBS and then lysed with 350-μl buffer RLT (Qiagen; containing β-mercaptoethanol at a concentration of 1:100), harvested and stored at −80°C for gene expression analysis. .. Cells were washed with RNAse-free PBS and then lysed with 350-μl buffer RLT (Qiagen; containing β-mercaptoethanol at a concentration of 1:100), harvested and stored at −80°C for gene expression analysis.

    Selection:

    Article Title: Allele-specific transcription factor binding to common and rare variants associated with disease and gene expression
    Article Snippet: The B cells were isolated from PBMCs by positive selection using CD19+ B-cell isolation kit (Miltenyi Biotec) according to manufacturer’s instructions. .. The harvested cells were stored in RLT buffer (QIAGEN) at −80 °C.

    Sample Prep:

    Article Title: Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies
    Article Snippet: To evaluate the LCM workflow and the number of sections required for collection of 100 ng of RNA we conducted a pilot study for RNA yield and quality. .. Following the described above approach to NanoString sample preparation we acquired 5μl lysates from pilot tissues, adjusted their volumes to 350μl with buffer RLT, then extracted RNA and qualified it as described above. .. Our LCM slide preparation, dissection and NanoString lysate preparation protocols effectively preserved RNA in the LCM targets ( ).

    In Vitro:

    Article Title: Does Caspase-6 Have a Role in Perinatal Brain Injury?
    Article Snippet: Paragraph title: In vitro Lipopolysaccharide and Q-VD-OPh Stimulation ... Cells were washed with RNAse-free PBS and then lysed with 350-μl buffer RLT (Qiagen; containing β-mercaptoethanol at a concentration of 1:100), harvested and stored at −80°C for gene expression analysis.

    Spectrophotometry:

    Article Title: Targeting Endothelial Adhesion Molecule Transcription for Treatment of Inflammatory Disease: A Proof-of-Concept Study
    Article Snippet: At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction. .. At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction.

    Article Title: Cardiac Non-myocyte Cells Show Enhanced Pharmacological Function Suggestive of Contractile Maturity in Stem Cell Derived Cardiomyocyte Microtissues
    Article Snippet: All primary cells were detached with prewarmed Accutase (Sigma, A6964) for 5 min at 37°C, 5% CO2 , centrifuged for 3 min at 1200 ×g before lysis in RLT buffer (Qiagen) and stored at −80 °C until processing. .. All primary cells were detached with prewarmed Accutase (Sigma, A6964) for 5 min at 37°C, 5% CO2 , centrifuged for 3 min at 1200 ×g before lysis in RLT buffer (Qiagen) and stored at −80 °C until processing.

    Sampling:

    Article Title: Pathogenic Differences between Nipah Virus Bangladesh and Malaysia Strains in Primates: Implications for Antibody Therapy
    Article Snippet: Immediately following sampling, 100 μl of blood was added to 600 μl of AVL viral lysis buffer (Qiagen) for RNA extraction. .. RNAlater was completely removed, and tissues were homogenized in 600 μl RLT buffer (Qiagen) in a 2-mL cryovial using a tissue lyser (Qiagen) and ceramic beads.

    Concentration Assay:

    Article Title: Allele-specific transcription factor binding to common and rare variants associated with disease and gene expression
    Article Snippet: The cells were stimulated with a phosphorothioate-modified CpG A oligonucleotide ODN2216 (CyberGene) at the concentration of 3 μg/ml and incubated for 5 h at 37 °C with 5 % CO2 . .. The harvested cells were stored in RLT buffer (QIAGEN) at −80 °C.

    Article Title: A virosomal respiratory syncytial virus vaccine adjuvanted with monophosphoryl lipid A provides protection against viral challenge without priming for enhanced disease in cotton rats) A virosomal respiratory syncytial virus vaccine adjuvanted with monophosphoryl lipid A provides protection against viral challenge without priming for enhanced disease in cotton rats
    Article Snippet: Lung samples stored in RNA later (Qiagen) were transferred to tubes containing 300 μl RLT buffer (Qiagen) and homogenized using disposable pestles with a pestle motor (VWR, Amsterdam, the Netherlands). .. Subsequently RNA was isolated with an RNeasy mini kit (Qiagen) according to the manufacturer's protocol.

    Article Title: Normalization with Corresponding Naïve Tissue Minimizes Bias Caused by Commercial Reverse Transcription Kits on Quantitative Real-Time PCR Results
    Article Snippet: For RNA extraction, brain tissue samples were lysed with RLT-buffer (Qiagen, Hilden, Germany) and homogenized with a MM300 mill mixer (Retsch, Haan, Germany) at 30 Hz for 2 minutes. .. RNA quality was confirmed via gel electrophoresis (Experion Automated Electrophoresis System, Bio-Rad, Munich, Germany) for RQI (RNA quality indicator) ≥8 [ ].

    Article Title: Targeting Endothelial Adhesion Molecule Transcription for Treatment of Inflammatory Disease: A Proof-of-Concept Study
    Article Snippet: At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction. .. At the conclusion of retinal endothelial cell manipulations, culture medium was replaced with Buffer RLT (Qiagen, Hilden, Germany) with 0.55 mM β -mercaptoethanol (Sigma-Aldrich, St. Louis, MO), and cells were frozen at −80°C prior to RNA extraction.

    Article Title: Does Caspase-6 Have a Role in Perinatal Brain Injury?
    Article Snippet: The entire supernatant was collected (1 ml) at 6, 24 or 48 h; 300 μl was used immediately for a lactate dehydrogenase assay (Roche, USA) and the remaining supernatant stored at −80°C until further analysis. .. Cells were washed with RNAse-free PBS and then lysed with 350-μl buffer RLT (Qiagen; containing β-mercaptoethanol at a concentration of 1:100), harvested and stored at −80°C for gene expression analysis. .. All time points were run in each independent experiment and all group experiments were performed a minimum of 3-6 independent times.

    Staining:

    Article Title: Genetic encoding of DNA nanostructures and their self-assembly in living bacteria
    Article Snippet: Cells were inoculated in 500 μl LB Miller Broth with antibiotics in a 96-well plate covered with a breathable membrane (AeraSeal, Excel Scientific) at 37 °C and 1,000 r.p.m. for 16 h. Overnight cultures were then diluted 1,000-fold by mixing 10 μl of the culture into 10 ml of LB Miller Broth containing the appropriate inducer and incubated at 37 °C and 250 r.p.m. for 18 h. After incubation, the DNA nanostructures were purified using the following protocol: (i) Cells were centrifuged at 5,000g for 7 min at 4 °C; (ii) the supernatant was removed; (iii) cells were resuspended in 200 μl of TE buffer (10 mM Tris-EDTA) containing 3 mg ml−1 of Lysozyme; (iv) 700 μl of RLT buffer (Qiagen, #79216) was added; (v) the resulting solution was centrifuged at 21,130 g for 2 min in order to remove the insoluble materials and the supernatant is transferred to a clean tube; (vi) 500 μl of 100% ethanol was added to the supernatant; (vii) 700 μl of the sample was transferred into a QIAquick Spin Column (Qiagen, #1018215) and centrifuged at 15,000g for 15 s; (viii) step vii was repeated until all the supernatant solution from step vi has passed through the same column tube (the flow through was discarded after each step); (ix) 700 μl of RW1 buffer (Qiagen, # 1053394) was added to the collection tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (x) 500 μl RPE buffer (Qiagen, # 1018013) was pipetted into the column tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (xi) 500 μl Buffer RPE (Mat. .. Cells were inoculated in 500 μl LB Miller Broth with antibiotics in a 96-well plate covered with a breathable membrane (AeraSeal, Excel Scientific) at 37 °C and 1,000 r.p.m. for 16 h. Overnight cultures were then diluted 1,000-fold by mixing 10 μl of the culture into 10 ml of LB Miller Broth containing the appropriate inducer and incubated at 37 °C and 250 r.p.m. for 18 h. After incubation, the DNA nanostructures were purified using the following protocol: (i) Cells were centrifuged at 5,000g for 7 min at 4 °C; (ii) the supernatant was removed; (iii) cells were resuspended in 200 μl of TE buffer (10 mM Tris-EDTA) containing 3 mg ml−1 of Lysozyme; (iv) 700 μl of RLT buffer (Qiagen, #79216) was added; (v) the resulting solution was centrifuged at 21,130 g for 2 min in order to remove the insoluble materials and the supernatant is transferred to a clean tube; (vi) 500 μl of 100% ethanol was added to the supernatant; (vii) 700 μl of the sample was transferred into a QIAquick Spin Column (Qiagen, #1018215) and centrifuged at 15,000g for 15 s; (viii) step vii was repeated until all the supernatant solution from step vi has passed through the same column tube (the flow through was discarded after each step); (ix) 700 μl of RW1 buffer (Qiagen, # 1053394) was added to the collection tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (x) 500 μl RPE buffer (Qiagen, # 1018013) was pipetted into the column tube and centrifuged for 15 s at 15,000g (the flow through was discarded); (xi) 500 μl Buffer RPE (Mat.

    Gradient Centrifugation:

    Article Title: Gene expression of INPP5F as an independent prognostic marker in fludarabine-based therapy of chronic lymphocytic leukemia
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were obtained by Ficoll density gradient centrifugation. .. CLL samples were positively enriched by magnetic cell sorting using CD19 microbeads and MACS columns (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and resuspended in RLT buffer (Qiagen, Crawley, West Sussex, UK).

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    Qiagen rlt lysis buffer provided in the rneasy kit qiagen hilden germany
    Rlt Lysis Buffer Provided In The Rneasy Kit Qiagen Hilden Germany, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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