rneasy minelute column  (Qiagen)


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    Name:
    RNeasy MinElute Cleanup Kit
    Description:
    For RNA cleanup and concentration with small elution volumes. Kit contents: Qiagen RNeasy MinElute Cleanup Kit, 50 preps, 200L Sample, 10 to 14L Elution Volume, Crude RNA preps Sample, Total RNA Purification, Silica Technology, MinElute Spin Column Format, Ideal for PCR, qPCR, Real-time PCR, Microarray, Includes 50 RNeasy MinElute Spin Columns, Collection Tubes (1.5mL and 2mL), RNase-free Reagents and Buffers. Benefits: Efficient cleanup of enzymatic reactions. Concentration of small amounts of RNA to only 10 µl. Cleanup of RNA purified by different methods. High-quality RNA in less than 15 m
    Catalog Number:
    74204
    Price:
    None
    Category:
    RNeasy MinElute Cleanup Kit
    Score:
    85
    Buy from Supplier


    Structured Review

    Qiagen rneasy minelute column
    RNeasy MinElute Cleanup Kit
    For RNA cleanup and concentration with small elution volumes. Kit contents: Qiagen RNeasy MinElute Cleanup Kit, 50 preps, 200L Sample, 10 to 14L Elution Volume, Crude RNA preps Sample, Total RNA Purification, Silica Technology, MinElute Spin Column Format, Ideal for PCR, qPCR, Real-time PCR, Microarray, Includes 50 RNeasy MinElute Spin Columns, Collection Tubes (1.5mL and 2mL), RNase-free Reagents and Buffers. Benefits: Efficient cleanup of enzymatic reactions. Concentration of small amounts of RNA to only 10 µl. Cleanup of RNA purified by different methods. High-quality RNA in less than 15 m
    https://www.bioz.com/result/rneasy minelute column/product/Qiagen
    Average 99 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    rneasy minelute column - by Bioz Stars, 2019-12
    99/100 stars

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    Related Articles

    Transduction:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: The cells were immediately cocultured with a GFP-2A-puromycin IDLV at a multiplicity of infection of 5 with subsequent spinning transduction. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    Clone Assay:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: Off-target analysis, deep sequencing, and KAT2B cloning. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: Paragraph title: Cloning, sequencing and RNA probe generation ... Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer.

    Centrifugation:

    Article Title: An In Vivo High-Throughput Screening Approach Targeting the Type IV Secretion System Component VirB8 Identified Inhibitors of Brucella abortus 2308 Proliferation
    Article Snippet: This solution was put onto a column, which was washed with 700 μl wash solution 1 and two times with 500 μl wash solution 2/3 by centrifugation. .. Before cDNA synthesis, the RNA was first treated twice with DNase and then cleaned up using Qiagen's RNeasy MinElute cleanup kit.

    Amplification:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: The high-throughput insertion site analysis pipeline was utilized for vector trimming and genome alignment, and IS/CLIS identification was determined. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles. .. PCR products were TA cloned into the pCR-4-TOPO Vector and Sanger sequenced with M13 Forward, M13 Reverse, and KAT2B1 internal: 5′-TACCTCGGTACGAAACCACACAGG-3′.

    Article Title: Interactions of the Hdm2/p53 and Proteasome Pathways May Enhance the Antitumor Activity of Bortezomib
    Article Snippet: RNA was extracted with TRIZOL-LS (Invitrogen), further treated with DNase I, and purified with the RNeasy MinElute Cleanup kit (Qiagen). .. Reverse transcription was accomplished with random hexamers and Superscript II, followed by incubation with RNase H (Invitrogen).

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: Digoxigenin (DIG)-labeled and fluorescein (FITC)-labeled antisense and sense RNAs were synthesized from purified pGEM-T easy plasmid DNA that was amplified using SP6 and T7 primers via PCR. .. Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer.

    Article Title: Inferior olivary TMEM16B mediates cerebellar motor learning
    Article Snippet: Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions. .. Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions.

    Article Title: Direct regulation of nucleosome density by the conserved AAA-ATPase Yta7
    Article Snippet: Total RNA was digested with DNase I (Roche) and purified using an RNeasy MinElute kit (Qiagen). .. Quantitative PCR (qPCR) on cDNA was performed using an MX3000P qPCR machine (Agilent) and a Dynamo HS SYBR Green qPCR kit (NEB).

    Mass Spectrometry:

    Article Title: Nucleolar GTP-Binding Protein 1-2 (NOG1-2) Interacts with Jasmonate-ZIMDomain Protein 9 (JAZ9) to Regulate Stomatal Aperture during Plant Immunity
    Article Snippet: Arabidopsis seedlings were grown for seven days on ½ MS in controlled conditions with a 16 h light, 8 h dark cycle at 24 °C. .. Total RNA from three biological replicates of nog1-2 , jaz9 , and Col-0 leaves were isolated and purified by using the RNeasy MinElute Cleanup Kit (Qiagen, Germantown, MD, USA) and used for two-channel microarray.

    Synthesized:

    Article Title: Environmental Nitrate Stimulates Abscisic Acid Accumulation in Arabidopsis Root Tips by Releasing It from Inactive Stores
    Article Snippet: Total RNA was isolated using a Qiagen RNeasy Plant Mini Kit (Qiagen) and subsequently treated with Turbo DNase-free (Ambion) to remove genomic DNA contamination, then cleaned and concentrated with a Qiagen RNeasy MinElute Cleanup Kit (Qiagen). .. Total RNA was isolated using a Qiagen RNeasy Plant Mini Kit (Qiagen) and subsequently treated with Turbo DNase-free (Ambion) to remove genomic DNA contamination, then cleaned and concentrated with a Qiagen RNeasy MinElute Cleanup Kit (Qiagen).

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: Digoxigenin (DIG)-labeled and fluorescein (FITC)-labeled antisense and sense RNAs were synthesized from purified pGEM-T easy plasmid DNA that was amplified using SP6 and T7 primers via PCR. .. Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer.

    Article Title: Direct and indirect parental exposure to endocrine disruptors and elevated temperature influences gene expression across generations in a euryhaline model fish
    Article Snippet: Samples containing trace amounts of genomic DNA were treated with DNase I (Invitrogen™, Waltham, MA) and cleaned up using RNeasy MinElute Cleanup Kit (Qiagen, Hilden, Germany). .. RNA concentrations were adjusted to a concentration of 100 ng/µl in 96-well plates, randomizing samples within each generation and tissue type prior to cDNA synthesis.

    Article Title: CFTR-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development
    Article Snippet: To prepare mRNA, the expression vector was first linearized by restriction endonuclease and then purified with PCR purification kit. .. The capped mRNA was synthesized in vitro using mMESSAGE mMACHINE SP6 kit (Ambion, Waltham, MA, USA, AM1340). mRNA was purified by RNA clean-up kit (Qiagen, Valencia, CA, USA, 74204). .. Xenopus embryos were injected at the two-cell stage two times with a mixture of 100 pg β -catenin/TCF reporter construct and 10 pg pRL-CMV (Renilla luciferase).

    Article Title: Transcriptional plasticity promotes primary and acquired resistance to BET inhibition
    Article Snippet: Mononuclear cells (MNC) were isolated using Ficoll and stored in liquid nitrogen until used. .. After thawing the viability of cells ranged from 75% to 85% as assessed by trypan blue exclusion.RNA was isolated from BM or peripheral blood MNC using the RNeasy MinElute CleanupKit (Qiagen). cDNA was synthesized using RQ1 DNase buffer, RQ1 DNase and RQ1 DNase Stop Solution (Promega), Moloney murine leukemia virus reverse transcriptase, random primers, First Strand buffer, dNTPs (100 mM), and RNasin (Invitrogen) according to the manufacturer’s instructions. .. MNC were cultured in 96-well plates (TPP) in RPMI-1640 medium (Lonza) with 10% fetal calf serum in the absence or presence of JQ1 (50–5,000 nM) at 37°C for 48 hours.

    Quantitative RT-PCR:

    Article Title: An In Vivo High-Throughput Screening Approach Targeting the Type IV Secretion System Component VirB8 Identified Inhibitors of Brucella abortus 2308 Proliferation
    Article Snippet: Paragraph title: Quantitative RT-PCR analysis. ... Before cDNA synthesis, the RNA was first treated twice with DNase and then cleaned up using Qiagen's RNeasy MinElute cleanup kit.

    Article Title: INTUSSUSCEPTIVE-LIKE ANGIOGENESIS IN HUMAN FETAL LUNG XENOGRAFTS: LINK WITH BRONCHOPULMONARY DYSPLASIA-ASSOCIATED MICROVASCULAR DYSANGIOGENESIS?
    Article Snippet: Paragraph title: Laser-capture microdissection, RNA extraction, microarray and qRT-PCR analysis of gene expression in xenografts ... RNA was further purified and concentrated using the RNEasy Minelute Cleanup kit (Qiagen, Valencia, CA) and then evaluated by the Agilent Bioanalyzer using an RNA 6000 Nano or Pico LabChip (Agilent Technologies, Santa Clara, CA).

    Article Title: Environmental Nitrate Stimulates Abscisic Acid Accumulation in Arabidopsis Root Tips by Releasing It from Inactive Stores
    Article Snippet: Paragraph title: RNA Extraction and Quantitative RT-PCR ... Total RNA was isolated using a Qiagen RNeasy Plant Mini Kit (Qiagen) and subsequently treated with Turbo DNase-free (Ambion) to remove genomic DNA contamination, then cleaned and concentrated with a Qiagen RNeasy MinElute Cleanup Kit (Qiagen).

    Article Title: Combining Genetic Perturbations and Proteomics to Examine Kinase-Phosphatase Networks in Drosophila Embryos
    Article Snippet: Paragraph title: RNA isolation, reverse transcription, and RT-qPCR ... Genomic DNA was eliminated by incubation with DNAse (Qiagen) and samples processed for cleanup with an RNeasy MinElute Cleanup Kit (Qiagen).

    Article Title: Direct regulation of nucleosome density by the conserved AAA-ATPase Yta7
    Article Snippet: Paragraph title: RNA Isolation and Quantitative RT-PCR. ... Total RNA was digested with DNase I (Roche) and purified using an RNeasy MinElute kit (Qiagen).

    Cytotoxicity Assay:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles. .. PCR products were TA cloned into the pCR-4-TOPO Vector and Sanger sequenced with M13 Forward, M13 Reverse, and KAT2B1 internal: 5′-TACCTCGGTACGAAACCACACAGG-3′.

    Real-time Polymerase Chain Reaction:

    Article Title: Environmental Nitrate Stimulates Abscisic Acid Accumulation in Arabidopsis Root Tips by Releasing It from Inactive Stores
    Article Snippet: Total RNA was isolated using a Qiagen RNeasy Plant Mini Kit (Qiagen) and subsequently treated with Turbo DNase-free (Ambion) to remove genomic DNA contamination, then cleaned and concentrated with a Qiagen RNeasy MinElute Cleanup Kit (Qiagen). .. The quality and yield of cDNA was assessed using a NanoDrop1000 spectrophotometer (Thermo Scientific).

    Article Title: Interactions of the Hdm2/p53 and Proteasome Pathways May Enhance the Antitumor Activity of Bortezomib
    Article Snippet: RNA was extracted with TRIZOL-LS (Invitrogen), further treated with DNase I, and purified with the RNeasy MinElute Cleanup kit (Qiagen). .. Reverse transcription was accomplished with random hexamers and Superscript II, followed by incubation with RNase H (Invitrogen).

    Article Title: Direct and indirect parental exposure to endocrine disruptors and elevated temperature influences gene expression across generations in a euryhaline model fish
    Article Snippet: Samples containing trace amounts of genomic DNA were treated with DNase I (Invitrogen™, Waltham, MA) and cleaned up using RNeasy MinElute Cleanup Kit (Qiagen, Hilden, Germany). .. Complimentary DNA (cDNA) was synthesized using Superscript III (Invitrogen, Carlsbad, CA, USA), following manufacture protocols and methods in .

    Article Title:
    Article Snippet: Paragraph title: Real-time Quantitative PCR ... RNA from sorted populations of cells was isolated using RNeasy MinElute Cleanup kit (Qiagen 74204).

    Article Title: Direct regulation of nucleosome density by the conserved AAA-ATPase Yta7
    Article Snippet: Total RNA was digested with DNase I (Roche) and purified using an RNeasy MinElute kit (Qiagen). .. Oligo(dT) priming was used for all experiments except those illustrated in – , for which random hexamer priming was used.

    Microarray:

    Article Title: INTUSSUSCEPTIVE-LIKE ANGIOGENESIS IN HUMAN FETAL LUNG XENOGRAFTS: LINK WITH BRONCHOPULMONARY DYSPLASIA-ASSOCIATED MICROVASCULAR DYSANGIOGENESIS?
    Article Snippet: Paragraph title: Laser-capture microdissection, RNA extraction, microarray and qRT-PCR analysis of gene expression in xenografts ... RNA was further purified and concentrated using the RNEasy Minelute Cleanup kit (Qiagen, Valencia, CA) and then evaluated by the Agilent Bioanalyzer using an RNA 6000 Nano or Pico LabChip (Agilent Technologies, Santa Clara, CA).

    Article Title: Nucleolar GTP-Binding Protein 1-2 (NOG1-2) Interacts with Jasmonate-ZIMDomain Protein 9 (JAZ9) to Regulate Stomatal Aperture during Plant Immunity
    Article Snippet: Arabidopsis seedlings were grown for seven days on ½ MS in controlled conditions with a 16 h light, 8 h dark cycle at 24 °C. .. Total RNA from three biological replicates of nog1-2 , jaz9 , and Col-0 leaves were isolated and purified by using the RNeasy MinElute Cleanup Kit (Qiagen, Germantown, MD, USA) and used for two-channel microarray. .. Experimental procedures of transcriptome analysis using Affymetrix ATH1 arrays were performed as described in the Affymetrix manual.

    Incubation:

    Article Title: Combining Genetic Perturbations and Proteomics to Examine Kinase-Phosphatase Networks in Drosophila Embryos
    Article Snippet: RNA was isolated by guanidinium thiocyanate-phenol-chloroform extraction using TRIzol (Life Technologies) and glass bead based cell disruption. .. Genomic DNA was eliminated by incubation with DNAse (Qiagen) and samples processed for cleanup with an RNeasy MinElute Cleanup Kit (Qiagen). .. One microgram of purified RNA was incubated with a mix of oligo (dT) and random hexamer primers and iScript reverse transcriptase (iScript cDNA Synthesis Kit, Bio-Rad) for cDNA synthesis. cDNA was used as template for amplification with validated primers in iQ SYBR Green Supermix with a CFX96 Real-Time PCR detection system (Bio-Rad).

    Article Title: Transcriptional plasticity promotes primary and acquired resistance to BET inhibition
    Article Snippet: After thawing the viability of cells ranged from 75% to 85% as assessed by trypan blue exclusion.RNA was isolated from BM or peripheral blood MNC using the RNeasy MinElute CleanupKit (Qiagen). cDNA was synthesized using RQ1 DNase buffer, RQ1 DNase and RQ1 DNase Stop Solution (Promega), Moloney murine leukemia virus reverse transcriptase, random primers, First Strand buffer, dNTPs (100 mM), and RNasin (Invitrogen) according to the manufacturer’s instructions. .. After thawing the viability of cells ranged from 75% to 85% as assessed by trypan blue exclusion.RNA was isolated from BM or peripheral blood MNC using the RNeasy MinElute CleanupKit (Qiagen). cDNA was synthesized using RQ1 DNase buffer, RQ1 DNase and RQ1 DNase Stop Solution (Promega), Moloney murine leukemia virus reverse transcriptase, random primers, First Strand buffer, dNTPs (100 mM), and RNasin (Invitrogen) according to the manufacturer’s instructions.

    Formalin-fixed Paraffin-Embedded:

    Article Title: INTUSSUSCEPTIVE-LIKE ANGIOGENESIS IN HUMAN FETAL LUNG XENOGRAFTS: LINK WITH BRONCHOPULMONARY DYSPLASIA-ASSOCIATED MICROVASCULAR DYSANGIOGENESIS?
    Article Snippet: Total RNA was extracted from the captured areas using the RecoverAll Total Nucleic Acid Extraction kits for formalin-fixed, paraffin-embedded (FFPE) tissues (Ambion, Austin, TX). .. RNA was further purified and concentrated using the RNEasy Minelute Cleanup kit (Qiagen, Valencia, CA) and then evaluated by the Agilent Bioanalyzer using an RNA 6000 Nano or Pico LabChip (Agilent Technologies, Santa Clara, CA).

    Infection:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: The cells were immediately cocultured with a GFP-2A-puromycin IDLV at a multiplicity of infection of 5 with subsequent spinning transduction. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    Article Title: Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii
    Article Snippet: Paragraph title: Bacterial RNA extraction from murine pneumonia infection ... Total RNA was immediately extracted from both samples using the RNeasy Mini Kit (Qiagen #74104), treated with DNAse I (Invitrogen, # 18068015) and purified with RNeasy MinElute Cleanup Kit (Qiagen, #74204).

    Expressing:

    Article Title: INTUSSUSCEPTIVE-LIKE ANGIOGENESIS IN HUMAN FETAL LUNG XENOGRAFTS: LINK WITH BRONCHOPULMONARY DYSPLASIA-ASSOCIATED MICROVASCULAR DYSANGIOGENESIS?
    Article Snippet: Paragraph title: Laser-capture microdissection, RNA extraction, microarray and qRT-PCR analysis of gene expression in xenografts ... RNA was further purified and concentrated using the RNEasy Minelute Cleanup kit (Qiagen, Valencia, CA) and then evaluated by the Agilent Bioanalyzer using an RNA 6000 Nano or Pico LabChip (Agilent Technologies, Santa Clara, CA).

    Article Title: A novel mutant Na+/HCO3− cotransporter NBCe1 in a case of compound‐heterozygous inheritance of proximal renal tubular acidosis
    Article Snippet: Paragraph title: NBCe1‐A expression in oocytes ... cRNA was generated from Not I linearized NBCe1‐A‐EGFP.pGH19 cDNA using a T7 mMessage mMachine transcription kit (Life Technologies, Grand Island, NY, USA) and purified using an RNeasy MinElute cleanup kit (Qiagen, Valencia, CA, USA).

    Article Title: Combining Genetic Perturbations and Proteomics to Examine Kinase-Phosphatase Networks in Drosophila Embryos
    Article Snippet: Genomic DNA was eliminated by incubation with DNAse (Qiagen) and samples processed for cleanup with an RNeasy MinElute Cleanup Kit (Qiagen). .. One microgram of purified RNA was incubated with a mix of oligo (dT) and random hexamer primers and iScript reverse transcriptase (iScript cDNA Synthesis Kit, Bio-Rad) for cDNA synthesis. cDNA was used as template for amplification with validated primers in iQ SYBR Green Supermix with a CFX96 Real-Time PCR detection system (Bio-Rad).

    Article Title: Nucleolar GTP-Binding Protein 1-2 (NOG1-2) Interacts with Jasmonate-ZIMDomain Protein 9 (JAZ9) to Regulate Stomatal Aperture during Plant Immunity
    Article Snippet: Total RNA from three biological replicates of nog1-2 , jaz9 , and Col-0 leaves were isolated and purified by using the RNeasy MinElute Cleanup Kit (Qiagen, Germantown, MD, USA) and used for two-channel microarray. .. Total RNA from three biological replicates of nog1-2 , jaz9 , and Col-0 leaves were isolated and purified by using the RNeasy MinElute Cleanup Kit (Qiagen, Germantown, MD, USA) and used for two-channel microarray.

    Article Title: Direct and indirect parental exposure to endocrine disruptors and elevated temperature influences gene expression across generations in a euryhaline model fish
    Article Snippet: Paragraph title: Gene expression quantification ... Samples containing trace amounts of genomic DNA were treated with DNase I (Invitrogen™, Waltham, MA) and cleaned up using RNeasy MinElute Cleanup Kit (Qiagen, Hilden, Germany).

    Article Title: CFTR-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development
    Article Snippet: To prepare mRNA, the expression vector was first linearized by restriction endonuclease and then purified with PCR purification kit. .. The capped mRNA was synthesized in vitro using mMESSAGE mMACHINE SP6 kit (Ambion, Waltham, MA, USA, AM1340). mRNA was purified by RNA clean-up kit (Qiagen, Valencia, CA, USA, 74204).

    Western Blot:

    Article Title: Inferior olivary TMEM16B mediates cerebellar motor learning
    Article Snippet: Paragraph title: RT-PCR and Western Blotting ... Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions.

    Over Expression:

    Article Title: CFTR-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development
    Article Snippet: Paragraph title: Knockdown and overexpression in Xenopus laevis ... The capped mRNA was synthesized in vitro using mMESSAGE mMACHINE SP6 kit (Ambion, Waltham, MA, USA, AM1340). mRNA was purified by RNA clean-up kit (Qiagen, Valencia, CA, USA, 74204).

    Derivative Assay:

    Article Title: INTUSSUSCEPTIVE-LIKE ANGIOGENESIS IN HUMAN FETAL LUNG XENOGRAFTS: LINK WITH BRONCHOPULMONARY DYSPLASIA-ASSOCIATED MICROVASCULAR DYSANGIOGENESIS?
    Article Snippet: Graft regions without PFA, derived from the same graft, served as control tissue for graft regions with PFA. .. RNA was further purified and concentrated using the RNEasy Minelute Cleanup kit (Qiagen, Valencia, CA) and then evaluated by the Agilent Bioanalyzer using an RNA 6000 Nano or Pico LabChip (Agilent Technologies, Santa Clara, CA).

    Gas Chromatography:

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: PCR amplification reactions were performed with Phusion HF DNA polymerase, Q5 high-fidelity DNA polymerase and Q5 high GC enhancer buffer (New England Biolabs, Ipswich, MA), according to the manufacturer’s suggested ratios. .. Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer.

    Cell Culture:

    Article Title: Interactions of the Hdm2/p53 and Proteasome Pathways May Enhance the Antitumor Activity of Bortezomib
    Article Snippet: MM.1S-GFP/luc cells were cultured for 24 h in the presence or absence of the BMSC line HS-5 and then sorted by fluorescence activated cell sorting (FACS) cell sorter (BD FACSAria cell sorter, Becton Dickinson). .. RNA was extracted with TRIZOL-LS (Invitrogen), further treated with DNase I, and purified with the RNeasy MinElute Cleanup kit (Qiagen).

    Article Title: Transcription Elongation Factor GreA Plays a Key Role in Cellular Invasion and Virulence of Francisella tularensis subsp. novicida
    Article Snippet: Overnight cultures of wild-type strain U112 and the Δ greA mutant were diluted 100-fold with fresh TSB and cultured in triplicate to mid-log phase on a shaker. .. The prepared RNAs were analyzed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and Agilent 2100 Bioanalyzer (Agilent, San Diego, CA, USA), and purified with the RNeasy MinElute Cleanup Kit (Qiagen).

    Generated:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: Forty-eight hours post gene transfer, 400 ng/ml of puromycin was added, and the cells were expanded for 7 additional days followed by genomic DNA isolation. nrLAM PCR or LAM PCR with MseI or MluCI , , was performed, and deep sequencing data were generated with the Illumina MiSeq platform (San Diego, CA). .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    Article Title: A novel mutant Na+/HCO3− cotransporter NBCe1 in a case of compound‐heterozygous inheritance of proximal renal tubular acidosis
    Article Snippet: OR3 medium contains 14 g L−1 of Leibovitz's L‐15 medium (Thermo Fisher Scientific), 100 units mL–1 penicillin, 100 μg mL−1 streptomycin and 5 m m Hepes, adjusted to pH 7.5 using NaOH, and then adjusted to 195 ± 5 mosmol kg−1 osmolality with H2 O. Osmolality was measured using a Vapro vapor pressure osmometer (Wescor, Logan, UT, USA). .. cRNA was generated from Not I linearized NBCe1‐A‐EGFP.pGH19 cDNA using a T7 mMessage mMachine transcription kit (Life Technologies, Grand Island, NY, USA) and purified using an RNeasy MinElute cleanup kit (Qiagen, Valencia, CA, USA). .. Except where specifically stated otherwise, de‐folliculated oocytes were injected with 25 nl of 1000 ng nl−1 cRNA or 25 nl of H2 O.

    Article Title: Antennal transcriptome of Manduca sexta
    Article Snippet: RNA extracted from four male and four female animals was unified and further purified by using the RNeasy MinElute Clean up Kit (Qiagen) following the manufacturer's protocol. .. RNA extracted from four male and four female animals was unified and further purified by using the RNeasy MinElute Clean up Kit (Qiagen) following the manufacturer's protocol.

    Polymerase Chain Reaction:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: The high-throughput insertion site analysis pipeline was utilized for vector trimming and genome alignment, and IS/CLIS identification was determined. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles. .. PCR products were TA cloned into the pCR-4-TOPO Vector and Sanger sequenced with M13 Forward, M13 Reverse, and KAT2B1 internal: 5′-TACCTCGGTACGAAACCACACAGG-3′.

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: Digoxigenin (DIG)-labeled and fluorescein (FITC)-labeled antisense and sense RNAs were synthesized from purified pGEM-T easy plasmid DNA that was amplified using SP6 and T7 primers via PCR. .. Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer.

    Article Title:
    Article Snippet: RNA from sorted populations of cells was isolated using RNeasy MinElute Cleanup kit (Qiagen 74204). .. RNA from sorted populations of cells was isolated using RNeasy MinElute Cleanup kit (Qiagen 74204).

    Article Title: CFTR-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development
    Article Snippet: To prepare mRNA, the expression vector was first linearized by restriction endonuclease and then purified with PCR purification kit. .. The capped mRNA was synthesized in vitro using mMESSAGE mMACHINE SP6 kit (Ambion, Waltham, MA, USA, AM1340). mRNA was purified by RNA clean-up kit (Qiagen, Valencia, CA, USA, 74204).

    Article Title: Inferior olivary TMEM16B mediates cerebellar motor learning
    Article Snippet: Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions. .. RT-PCR was performed using the Platinum® Pfx DNA Polymerase (Life Technologies) with primers specific to TMEM16B and β-actin as described before ( ).

    Sequencing:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: Forty-eight hours post gene transfer, 400 ng/ml of puromycin was added, and the cells were expanded for 7 additional days followed by genomic DNA isolation. nrLAM PCR or LAM PCR with MseI or MluCI , , was performed, and deep sequencing data were generated with the Illumina MiSeq platform (San Diego, CA). .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: Paragraph title: Cloning, sequencing and RNA probe generation ... Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer.

    Article Title: Transcription Elongation Factor GreA Plays a Key Role in Cellular Invasion and Virulence of Francisella tularensis subsp. novicida
    Article Snippet: The prepared RNAs were analyzed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and Agilent 2100 Bioanalyzer (Agilent, San Diego, CA, USA), and purified with the RNeasy MinElute Cleanup Kit (Qiagen). .. The prepared RNAs were analyzed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and Agilent 2100 Bioanalyzer (Agilent, San Diego, CA, USA), and purified with the RNeasy MinElute Cleanup Kit (Qiagen).

    Article Title: CFTR-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development
    Article Snippet: In the loss-of-function analysis, antisense MO oligonucleotide against Xenopus CFTR RNA containing the sequence 5′-CTTTCTCCAGCGGCGTCTTCTGCAT-3′ (CFTR MO) or a standard control MO oligo containing the sequence 5′-CCTCTTACCTCAGTTACAATTTATA-3′ (control) was injected into the embryos at two-cell stage. .. The capped mRNA was synthesized in vitro using mMESSAGE mMACHINE SP6 kit (Ambion, Waltham, MA, USA, AM1340). mRNA was purified by RNA clean-up kit (Qiagen, Valencia, CA, USA, 74204).

    Article Title: Antennal transcriptome of Manduca sexta
    Article Snippet: Paragraph title: Expressed Sequence Tag Generation. ... RNA extracted from four male and four female animals was unified and further purified by using the RNeasy MinElute Clean up Kit (Qiagen) following the manufacturer's protocol.

    Article Title: Inferior olivary TMEM16B mediates cerebellar motor learning
    Article Snippet: Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions. .. Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions.

    Article Title: Persistent activation of interlinked type 2 airway epithelial gene networks in sputum-derived cells from aeroallergen-sensitized symptomatic asthmatics
    Article Snippet: Total RNA was extracted from good quality sputum (cell viability > 48%; squamous cell contamination < 32%; RNA integrity number > 6) employing TRIzol (Ambion) followed by RNeasy MinElute (QIAgen). .. Total RNA was extracted from good quality sputum (cell viability > 48%; squamous cell contamination < 32%; RNA integrity number > 6) employing TRIzol (Ambion) followed by RNeasy MinElute (QIAgen).

    Injection:

    Article Title: A novel mutant Na+/HCO3− cotransporter NBCe1 in a case of compound‐heterozygous inheritance of proximal renal tubular acidosis
    Article Snippet: cRNA was generated from Not I linearized NBCe1‐A‐EGFP.pGH19 cDNA using a T7 mMessage mMachine transcription kit (Life Technologies, Grand Island, NY, USA) and purified using an RNeasy MinElute cleanup kit (Qiagen, Valencia, CA, USA). .. Except where specifically stated otherwise, de‐folliculated oocytes were injected with 25 nl of 1000 ng nl−1 cRNA or 25 nl of H2 O.

    Article Title: CFTR-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development
    Article Snippet: In the loss-of-function analysis, antisense MO oligonucleotide against Xenopus CFTR RNA containing the sequence 5′-CTTTCTCCAGCGGCGTCTTCTGCAT-3′ (CFTR MO) or a standard control MO oligo containing the sequence 5′-CCTCTTACCTCAGTTACAATTTATA-3′ (control) was injected into the embryos at two-cell stage. .. The capped mRNA was synthesized in vitro using mMESSAGE mMACHINE SP6 kit (Ambion, Waltham, MA, USA, AM1340). mRNA was purified by RNA clean-up kit (Qiagen, Valencia, CA, USA, 74204).

    Recombinant:

    Article Title: Transcriptional plasticity promotes primary and acquired resistance to BET inhibition
    Article Snippet: After thawing the viability of cells ranged from 75% to 85% as assessed by trypan blue exclusion.RNA was isolated from BM or peripheral blood MNC using the RNeasy MinElute CleanupKit (Qiagen). cDNA was synthesized using RQ1 DNase buffer, RQ1 DNase and RQ1 DNase Stop Solution (Promega), Moloney murine leukemia virus reverse transcriptase, random primers, First Strand buffer, dNTPs (100 mM), and RNasin (Invitrogen) according to the manufacturer’s instructions. .. MNC were cultured in 96-well plates (TPP) in RPMI-1640 medium (Lonza) with 10% fetal calf serum in the absence or presence of JQ1 (50–5,000 nM) at 37°C for 48 hours.

    DNA Extraction:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: Forty-eight hours post gene transfer, 400 ng/ml of puromycin was added, and the cells were expanded for 7 additional days followed by genomic DNA isolation. nrLAM PCR or LAM PCR with MseI or MluCI , , was performed, and deep sequencing data were generated with the Illumina MiSeq platform (San Diego, CA). .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    In Vivo:

    Article Title: Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii
    Article Snippet: Then, a bronchoalveolar lavage (BAL) was performed to obtained bacteria suitable for RNA extraction ( in vivo samples). .. Total RNA was immediately extracted from both samples using the RNeasy Mini Kit (Qiagen #74104), treated with DNAse I (Invitrogen, # 18068015) and purified with RNeasy MinElute Cleanup Kit (Qiagen, #74204).

    RNA Sequencing Assay:

    Article Title: Biomarkers from circulating neutrophil transcriptomes have potential to detect unruptured intracranial aneurysms
    Article Snippet: RNA was purified using the RNeasy MinElute Cleanup Kit (Qiagen, Venlo, Limburg, Netherlands) and suspended in RNase-free water. .. The quality of the RNA samples was measured with an Agilent 2100 BioAnalyzer RNA 6000 Pico Chip (Agilent, Las Vegas, NV).

    Article Title: Transcription Elongation Factor GreA Plays a Key Role in Cellular Invasion and Virulence of Francisella tularensis subsp. novicida
    Article Snippet: Paragraph title: RNA-seq ... The prepared RNAs were analyzed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and Agilent 2100 Bioanalyzer (Agilent, San Diego, CA, USA), and purified with the RNeasy MinElute Cleanup Kit (Qiagen).

    Article Title: Persistent activation of interlinked type 2 airway epithelial gene networks in sputum-derived cells from aeroallergen-sensitized symptomatic asthmatics
    Article Snippet: Paragraph title: Transcriptome profiling by RNA-Seq ... Total RNA was extracted from good quality sputum (cell viability > 48%; squamous cell contamination < 32%; RNA integrity number > 6) employing TRIzol (Ambion) followed by RNeasy MinElute (QIAgen).

    Fluorescence:

    Article Title: Interactions of the Hdm2/p53 and Proteasome Pathways May Enhance the Antitumor Activity of Bortezomib
    Article Snippet: MM.1S-GFP/luc cells were cultured for 24 h in the presence or absence of the BMSC line HS-5 and then sorted by fluorescence activated cell sorting (FACS) cell sorter (BD FACSAria cell sorter, Becton Dickinson). .. RNA was extracted with TRIZOL-LS (Invitrogen), further treated with DNase I, and purified with the RNeasy MinElute Cleanup kit (Qiagen).

    Mutagenesis:

    Article Title: Transcription Elongation Factor GreA Plays a Key Role in Cellular Invasion and Virulence of Francisella tularensis subsp. novicida
    Article Snippet: Overnight cultures of wild-type strain U112 and the Δ greA mutant were diluted 100-fold with fresh TSB and cultured in triplicate to mid-log phase on a shaker. .. The prepared RNAs were analyzed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and Agilent 2100 Bioanalyzer (Agilent, San Diego, CA, USA), and purified with the RNeasy MinElute Cleanup Kit (Qiagen).

    Isolation:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: The high-throughput insertion site analysis pipeline was utilized for vector trimming and genome alignment, and IS/CLIS identification was determined. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles. .. PCR products were TA cloned into the pCR-4-TOPO Vector and Sanger sequenced with M13 Forward, M13 Reverse, and KAT2B1 internal: 5′-TACCTCGGTACGAAACCACACAGG-3′.

    Article Title: Environmental Nitrate Stimulates Abscisic Acid Accumulation in Arabidopsis Root Tips by Releasing It from Inactive Stores
    Article Snippet: Seedlings were grown as described above, after which either whole seedlings or roots were harvested for RNA isolation. .. Total RNA was isolated using a Qiagen RNeasy Plant Mini Kit (Qiagen) and subsequently treated with Turbo DNase-free (Ambion) to remove genomic DNA contamination, then cleaned and concentrated with a Qiagen RNeasy MinElute Cleanup Kit (Qiagen). .. All RNA samples were checked on a 2100 Bioanalyzer for quality and integrity.

    Article Title: Combining Genetic Perturbations and Proteomics to Examine Kinase-Phosphatase Networks in Drosophila Embryos
    Article Snippet: Paragraph title: RNA isolation, reverse transcription, and RT-qPCR ... Genomic DNA was eliminated by incubation with DNAse (Qiagen) and samples processed for cleanup with an RNeasy MinElute Cleanup Kit (Qiagen).

    Article Title: Nucleolar GTP-Binding Protein 1-2 (NOG1-2) Interacts with Jasmonate-ZIMDomain Protein 9 (JAZ9) to Regulate Stomatal Aperture during Plant Immunity
    Article Snippet: Arabidopsis seedlings were grown for seven days on ½ MS in controlled conditions with a 16 h light, 8 h dark cycle at 24 °C. .. Total RNA from three biological replicates of nog1-2 , jaz9 , and Col-0 leaves were isolated and purified by using the RNeasy MinElute Cleanup Kit (Qiagen, Germantown, MD, USA) and used for two-channel microarray. .. Experimental procedures of transcriptome analysis using Affymetrix ATH1 arrays were performed as described in the Affymetrix manual.

    Article Title:
    Article Snippet: Excess cells were removed in a large volume of PBS and the arrays were fixed in 4% paraformaldehyde solution for 10 min. Arrays were imaged using automated, high content fluorescence microscopy (IXmicro, MDC). .. RNA from sorted populations of cells was isolated using RNeasy MinElute Cleanup kit (Qiagen 74204). .. DNA was eliminated using TURBO DNA-free kit (Ambion AM1907) and the mRNA was converted to cDNA using high-capacity cDNA Reverse Transcription kit (Ambion 4374967).

    Article Title: Direct regulation of nucleosome density by the conserved AAA-ATPase Yta7
    Article Snippet: Paragraph title: RNA Isolation and Quantitative RT-PCR. ... Total RNA was digested with DNase I (Roche) and purified using an RNeasy MinElute kit (Qiagen).

    Article Title: Transcriptional plasticity promotes primary and acquired resistance to BET inhibition
    Article Snippet: Mononuclear cells (MNC) were isolated using Ficoll and stored in liquid nitrogen until used. .. After thawing the viability of cells ranged from 75% to 85% as assessed by trypan blue exclusion.RNA was isolated from BM or peripheral blood MNC using the RNeasy MinElute CleanupKit (Qiagen). cDNA was synthesized using RQ1 DNase buffer, RQ1 DNase and RQ1 DNase Stop Solution (Promega), Moloney murine leukemia virus reverse transcriptase, random primers, First Strand buffer, dNTPs (100 mM), and RNasin (Invitrogen) according to the manufacturer’s instructions. .. MNC were cultured in 96-well plates (TPP) in RPMI-1640 medium (Lonza) with 10% fetal calf serum in the absence or presence of JQ1 (50–5,000 nM) at 37°C for 48 hours.

    Labeling:

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: Template DNA was used with SP6 and T7 RNA polymerase (Invitrogen, Carlsbad, CA, USA), DIG and fluorescein labeling mix (Roche, Indianapolis, IN, USA) to generate probes. .. Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer.

    Purification:

    Article Title: INTUSSUSCEPTIVE-LIKE ANGIOGENESIS IN HUMAN FETAL LUNG XENOGRAFTS: LINK WITH BRONCHOPULMONARY DYSPLASIA-ASSOCIATED MICROVASCULAR DYSANGIOGENESIS?
    Article Snippet: Total RNA was extracted from the captured areas using the RecoverAll Total Nucleic Acid Extraction kits for formalin-fixed, paraffin-embedded (FFPE) tissues (Ambion, Austin, TX). .. RNA was further purified and concentrated using the RNEasy Minelute Cleanup kit (Qiagen, Valencia, CA) and then evaluated by the Agilent Bioanalyzer using an RNA 6000 Nano or Pico LabChip (Agilent Technologies, Santa Clara, CA). .. Total RNA (150 ng) extracted from FFPE blocks was used with the Affymetrix SensationPlus™FFPE Amplification and WT labeling Kit (cat # 902042) according to the manufacturer’s instructions.

    Article Title: A novel mutant Na+/HCO3− cotransporter NBCe1 in a case of compound‐heterozygous inheritance of proximal renal tubular acidosis
    Article Snippet: OR3 medium contains 14 g L−1 of Leibovitz's L‐15 medium (Thermo Fisher Scientific), 100 units mL–1 penicillin, 100 μg mL−1 streptomycin and 5 m m Hepes, adjusted to pH 7.5 using NaOH, and then adjusted to 195 ± 5 mosmol kg−1 osmolality with H2 O. Osmolality was measured using a Vapro vapor pressure osmometer (Wescor, Logan, UT, USA). .. cRNA was generated from Not I linearized NBCe1‐A‐EGFP.pGH19 cDNA using a T7 mMessage mMachine transcription kit (Life Technologies, Grand Island, NY, USA) and purified using an RNeasy MinElute cleanup kit (Qiagen, Valencia, CA, USA). .. Except where specifically stated otherwise, de‐folliculated oocytes were injected with 25 nl of 1000 ng nl−1 cRNA or 25 nl of H2 O.

    Article Title: Interactions of the Hdm2/p53 and Proteasome Pathways May Enhance the Antitumor Activity of Bortezomib
    Article Snippet: MM.1S-GFP/luc cells were cultured for 24 h in the presence or absence of the BMSC line HS-5 and then sorted by fluorescence activated cell sorting (FACS) cell sorter (BD FACSAria cell sorter, Becton Dickinson). .. RNA was extracted with TRIZOL-LS (Invitrogen), further treated with DNase I, and purified with the RNeasy MinElute Cleanup kit (Qiagen). .. Reverse transcription was accomplished with random hexamers and Superscript II, followed by incubation with RNase H (Invitrogen).

    Article Title: Nucleolar GTP-Binding Protein 1-2 (NOG1-2) Interacts with Jasmonate-ZIMDomain Protein 9 (JAZ9) to Regulate Stomatal Aperture during Plant Immunity
    Article Snippet: Arabidopsis seedlings were grown for seven days on ½ MS in controlled conditions with a 16 h light, 8 h dark cycle at 24 °C. .. Total RNA from three biological replicates of nog1-2 , jaz9 , and Col-0 leaves were isolated and purified by using the RNeasy MinElute Cleanup Kit (Qiagen, Germantown, MD, USA) and used for two-channel microarray. .. Experimental procedures of transcriptome analysis using Affymetrix ATH1 arrays were performed as described in the Affymetrix manual.

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: Template DNA was used with SP6 and T7 RNA polymerase (Invitrogen, Carlsbad, CA, USA), DIG and fluorescein labeling mix (Roche, Indianapolis, IN, USA) to generate probes. .. Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer. .. The in situ hybridization protocol for single-gene labeling was modified from Finnerty et al. [ ] and Perry et al. [ ].

    Article Title: Transcription Elongation Factor GreA Plays a Key Role in Cellular Invasion and Virulence of Francisella tularensis subsp. novicida
    Article Snippet: Total RNA of each sample was extracted using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA)/RNeasy Mini Kit (Qiagen, Dusseldorf, Germany). .. The prepared RNAs were analyzed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and Agilent 2100 Bioanalyzer (Agilent, San Diego, CA, USA), and purified with the RNeasy MinElute Cleanup Kit (Qiagen). .. The Ribo-Zero™ Magnetic Kit (Epicentre, Madison, WI, USA) was used to remove the rRNA, and the rRNA-depleted RNA was used to generate cDNA libraries with the NEBNext Ultra™ Directional RNA Library Prep Kit for Illumina® (NEB).

    Article Title: Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii
    Article Snippet: RNA extracted from bacteria grown in LB-flasks (OD600 = 1.0) at 37°C and 180 rpm was used as experimental control ( in vitro samples). .. Total RNA was immediately extracted from both samples using the RNeasy Mini Kit (Qiagen #74104), treated with DNAse I (Invitrogen, # 18068015) and purified with RNeasy MinElute Cleanup Kit (Qiagen, #74204). .. Final concentrations and purity grades of the samples were determined using a BioDrop μLITE (Isogen Life Science) and a Bioanalyzer 2100 (Agilent Technologies Inc.).

    Article Title: CFTR-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development
    Article Snippet: To prepare mRNA, the expression vector was first linearized by restriction endonuclease and then purified with PCR purification kit. .. The capped mRNA was synthesized in vitro using mMESSAGE mMACHINE SP6 kit (Ambion, Waltham, MA, USA, AM1340). mRNA was purified by RNA clean-up kit (Qiagen, Valencia, CA, USA, 74204). .. Xenopus embryos were injected at the two-cell stage two times with a mixture of 100 pg β -catenin/TCF reporter construct and 10 pg pRL-CMV (Renilla luciferase).

    Article Title: Antennal transcriptome of Manduca sexta
    Article Snippet: Total RNA was extracted by using the TRIzol method. .. RNA extracted from four male and four female animals was unified and further purified by using the RNeasy MinElute Clean up Kit (Qiagen) following the manufacturer's protocol. .. To prevent overrepresentation of the most common transcripts, the reverse-transcribed mRNAs converted to double-stranded cDNAs (see below) were normalized by using the duplex-specific nuclease method ( ).

    Article Title: Inferior olivary TMEM16B mediates cerebellar motor learning
    Article Snippet: For RT-PCR, retina, olfactory bulbs, hippocampi, and medulla oblongata were dissected from TMEM16B knockout mice and wild type controls (1.5 – 2 months old). .. Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions. .. Two micrograms of each total RNA were reverse transcribed into cDNA using the SuperScript® III First-Strand Synthesis System (Life Technologies).

    Article Title: Direct regulation of nucleosome density by the conserved AAA-ATPase Yta7
    Article Snippet: RNA isolation for all experiments except those illustrated in and and and was performed using the hot-phenol method ( ). .. Total RNA was digested with DNase I (Roche) and purified using an RNeasy MinElute kit (Qiagen). .. RNA for the experiments illustrated in and and and was isolated directly from whole-cell extract using an RNeasy Mini kit (Qiagen) with on-column DNase I (Qiagen) digestion. cDNA was synthesized using a Super-Script III First-Strand Synthesis System (Invitrogen).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Inferior olivary TMEM16B mediates cerebellar motor learning
    Article Snippet: Paragraph title: RT-PCR and Western Blotting ... Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions.

    Spectrophotometry:

    Article Title: Environmental Nitrate Stimulates Abscisic Acid Accumulation in Arabidopsis Root Tips by Releasing It from Inactive Stores
    Article Snippet: Total RNA was isolated using a Qiagen RNeasy Plant Mini Kit (Qiagen) and subsequently treated with Turbo DNase-free (Ambion) to remove genomic DNA contamination, then cleaned and concentrated with a Qiagen RNeasy MinElute Cleanup Kit (Qiagen). .. First-strand cDNA was synthesized from 1 µg of total RNA using an iScript kit (Bio-Rad).

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: Template DNA was used with SP6 and T7 RNA polymerase (Invitrogen, Carlsbad, CA, USA), DIG and fluorescein labeling mix (Roche, Indianapolis, IN, USA) to generate probes. .. Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer. .. The in situ hybridization protocol for single-gene labeling was modified from Finnerty et al. [ ] and Perry et al. [ ].

    Article Title: Transcription Elongation Factor GreA Plays a Key Role in Cellular Invasion and Virulence of Francisella tularensis subsp. novicida
    Article Snippet: Total RNA of each sample was extracted using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA)/RNeasy Mini Kit (Qiagen, Dusseldorf, Germany). .. The prepared RNAs were analyzed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and Agilent 2100 Bioanalyzer (Agilent, San Diego, CA, USA), and purified with the RNeasy MinElute Cleanup Kit (Qiagen). .. The Ribo-Zero™ Magnetic Kit (Epicentre, Madison, WI, USA) was used to remove the rRNA, and the rRNA-depleted RNA was used to generate cDNA libraries with the NEBNext Ultra™ Directional RNA Library Prep Kit for Illumina® (NEB).

    Article Title: Direct and indirect parental exposure to endocrine disruptors and elevated temperature influences gene expression across generations in a euryhaline model fish
    Article Snippet: RNA was extracted using RNeasy Mini Kits (Qiagen, Hilden, Germany) Total RNA concentration and 260/280 ratios (acceptable values > 1.6) were quantified using a Nanodrop 1000 spectrophotometer (ThermoFisher Scientific, Waltham, MA). .. Samples containing trace amounts of genomic DNA were treated with DNase I (Invitrogen™, Waltham, MA) and cleaned up using RNeasy MinElute Cleanup Kit (Qiagen, Hilden, Germany).

    CRISPR:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: Six samples for CRISPR/Cas9 and MT and four for TALEN were generated for OT analysis by electroporating Jurkat cells with 1 μg of MT or TALEN mRNA or 500 ng each of Cas9 mRNA and gRNA plasmid. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    cDNA Library Assay:

    Article Title: Antennal transcriptome of Manduca sexta
    Article Snippet: RNA extracted from four male and four female animals was unified and further purified by using the RNeasy MinElute Clean up Kit (Qiagen) following the manufacturer's protocol. .. RNA extracted from four male and four female animals was unified and further purified by using the RNeasy MinElute Clean up Kit (Qiagen) following the manufacturer's protocol.

    Mouse Assay:

    Article Title: Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii
    Article Snippet: All mice were maintained in the specific pathogen-free facility at the Technology Training Center of the Hospital of A Coruña (CHUAC, Spain). .. Total RNA was immediately extracted from both samples using the RNeasy Mini Kit (Qiagen #74104), treated with DNAse I (Invitrogen, # 18068015) and purified with RNeasy MinElute Cleanup Kit (Qiagen, #74204).

    Article Title: Inferior olivary TMEM16B mediates cerebellar motor learning
    Article Snippet: For RT-PCR, retina, olfactory bulbs, hippocampi, and medulla oblongata were dissected from TMEM16B knockout mice and wild type controls (1.5 – 2 months old). .. Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions.

    Chromatin Immunoprecipitation:

    Article Title: Biomarkers from circulating neutrophil transcriptomes have potential to detect unruptured intracranial aneurysms
    Article Snippet: RNA was purified using the RNeasy MinElute Cleanup Kit (Qiagen, Venlo, Limburg, Netherlands) and suspended in RNase-free water. .. Precise RNA concentration was measured at the core facility via the Quant-iT RiboGreen Assay (Invitrogen, Carlsbad, CA) with a TBS-380 Fluorometer (Promega, Madison, WI).

    Plasmid Preparation:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: The high-throughput insertion site analysis pipeline was utilized for vector trimming and genome alignment, and IS/CLIS identification was determined. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    Article Title: Molecular, phylogenetic and developmental analyses of Sall proteins in bilaterians
    Article Snippet: Digoxigenin (DIG)-labeled and fluorescein (FITC)-labeled antisense and sense RNAs were synthesized from purified pGEM-T easy plasmid DNA that was amplified using SP6 and T7 primers via PCR. .. Reactions were purified with RNeasy MinElute Cleanup kit (Qiagen, Valencia, CA), and probe concentrations were verified using a NanoDrop ND-1000 Spectrophotometer.

    Article Title: CFTR-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development
    Article Snippet: To prepare mRNA, the expression vector was first linearized by restriction endonuclease and then purified with PCR purification kit. .. The capped mRNA was synthesized in vitro using mMESSAGE mMACHINE SP6 kit (Ambion, Waltham, MA, USA, AM1340). mRNA was purified by RNA clean-up kit (Qiagen, Valencia, CA, USA, 74204).

    Software:

    Article Title: Interactions of the Hdm2/p53 and Proteasome Pathways May Enhance the Antitumor Activity of Bortezomib
    Article Snippet: RNA was extracted with TRIZOL-LS (Invitrogen), further treated with DNase I, and purified with the RNeasy MinElute Cleanup kit (Qiagen). .. Amplification reactions (25 μL, 100 ng cDNA/reaction) with Taqman FAM/MGB probes were done in a 7500 Real-time PCR System (Applied Biosystems) for select p53-regulated transcripts.

    Article Title: Transcription Elongation Factor GreA Plays a Key Role in Cellular Invasion and Virulence of Francisella tularensis subsp. novicida
    Article Snippet: The prepared RNAs were analyzed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and Agilent 2100 Bioanalyzer (Agilent, San Diego, CA, USA), and purified with the RNeasy MinElute Cleanup Kit (Qiagen). .. The prepared RNAs were analyzed with a NanoDrop 2000 spectrophotometer (Thermo Scientific) and Agilent 2100 Bioanalyzer (Agilent, San Diego, CA, USA), and purified with the RNeasy MinElute Cleanup Kit (Qiagen).

    SYBR Green Assay:

    Article Title: Environmental Nitrate Stimulates Abscisic Acid Accumulation in Arabidopsis Root Tips by Releasing It from Inactive Stores
    Article Snippet: Total RNA was isolated using a Qiagen RNeasy Plant Mini Kit (Qiagen) and subsequently treated with Turbo DNase-free (Ambion) to remove genomic DNA contamination, then cleaned and concentrated with a Qiagen RNeasy MinElute Cleanup Kit (Qiagen). .. The quality and yield of cDNA was assessed using a NanoDrop1000 spectrophotometer (Thermo Scientific).

    Article Title:
    Article Snippet: RNA from sorted populations of cells was isolated using RNeasy MinElute Cleanup kit (Qiagen 74204). .. RNA from sorted populations of cells was isolated using RNeasy MinElute Cleanup kit (Qiagen 74204).

    Article Title: Direct regulation of nucleosome density by the conserved AAA-ATPase Yta7
    Article Snippet: Total RNA was digested with DNase I (Roche) and purified using an RNeasy MinElute kit (Qiagen). .. Oligo(dT) priming was used for all experiments except those illustrated in – , for which random hexamer priming was used.

    RNA Extraction:

    Article Title: INTUSSUSCEPTIVE-LIKE ANGIOGENESIS IN HUMAN FETAL LUNG XENOGRAFTS: LINK WITH BRONCHOPULMONARY DYSPLASIA-ASSOCIATED MICROVASCULAR DYSANGIOGENESIS?
    Article Snippet: Paragraph title: Laser-capture microdissection, RNA extraction, microarray and qRT-PCR analysis of gene expression in xenografts ... RNA was further purified and concentrated using the RNEasy Minelute Cleanup kit (Qiagen, Valencia, CA) and then evaluated by the Agilent Bioanalyzer using an RNA 6000 Nano or Pico LabChip (Agilent Technologies, Santa Clara, CA).

    Article Title: Environmental Nitrate Stimulates Abscisic Acid Accumulation in Arabidopsis Root Tips by Releasing It from Inactive Stores
    Article Snippet: Paragraph title: RNA Extraction and Quantitative RT-PCR ... Total RNA was isolated using a Qiagen RNeasy Plant Mini Kit (Qiagen) and subsequently treated with Turbo DNase-free (Ambion) to remove genomic DNA contamination, then cleaned and concentrated with a Qiagen RNeasy MinElute Cleanup Kit (Qiagen).

    Article Title: Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii
    Article Snippet: Paragraph title: Bacterial RNA extraction from murine pneumonia infection ... Total RNA was immediately extracted from both samples using the RNeasy Mini Kit (Qiagen #74104), treated with DNAse I (Invitrogen, # 18068015) and purified with RNeasy MinElute Cleanup Kit (Qiagen, #74204).

    Agarose Gel Electrophoresis:

    Article Title: Direct and indirect parental exposure to endocrine disruptors and elevated temperature influences gene expression across generations in a euryhaline model fish
    Article Snippet: RNA purity was assessed via electrophoresis on a 1% agarose gel and visualized on a Gel Doc™ XR + Gel Documentation system (BioRad, Hercules, CA, USA). .. Samples containing trace amounts of genomic DNA were treated with DNase I (Invitrogen™, Waltham, MA) and cleaned up using RNeasy MinElute Cleanup Kit (Qiagen, Hilden, Germany).

    In Vitro:

    Article Title: INTUSSUSCEPTIVE-LIKE ANGIOGENESIS IN HUMAN FETAL LUNG XENOGRAFTS: LINK WITH BRONCHOPULMONARY DYSPLASIA-ASSOCIATED MICROVASCULAR DYSANGIOGENESIS?
    Article Snippet: RNA was further purified and concentrated using the RNEasy Minelute Cleanup kit (Qiagen, Valencia, CA) and then evaluated by the Agilent Bioanalyzer using an RNA 6000 Nano or Pico LabChip (Agilent Technologies, Santa Clara, CA). .. Total RNA (150 ng) extracted from FFPE blocks was used with the Affymetrix SensationPlus™FFPE Amplification and WT labeling Kit (cat # 902042) according to the manufacturer’s instructions.

    Article Title: Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii
    Article Snippet: RNA extracted from bacteria grown in LB-flasks (OD600 = 1.0) at 37°C and 180 rpm was used as experimental control ( in vitro samples). .. Total RNA was immediately extracted from both samples using the RNeasy Mini Kit (Qiagen #74104), treated with DNAse I (Invitrogen, # 18068015) and purified with RNeasy MinElute Cleanup Kit (Qiagen, #74204).

    Article Title: CFTR-β-catenin interaction regulates mouse embryonic stem cell differentiation and embryonic development
    Article Snippet: To prepare mRNA, the expression vector was first linearized by restriction endonuclease and then purified with PCR purification kit. .. The capped mRNA was synthesized in vitro using mMESSAGE mMACHINE SP6 kit (Ambion, Waltham, MA, USA, AM1340). mRNA was purified by RNA clean-up kit (Qiagen, Valencia, CA, USA, 74204). .. Xenopus embryos were injected at the two-cell stage two times with a mixture of 100 pg β -catenin/TCF reporter construct and 10 pg pRL-CMV (Renilla luciferase).

    Electrophoresis:

    Article Title: Direct and indirect parental exposure to endocrine disruptors and elevated temperature influences gene expression across generations in a euryhaline model fish
    Article Snippet: RNA purity was assessed via electrophoresis on a 1% agarose gel and visualized on a Gel Doc™ XR + Gel Documentation system (BioRad, Hercules, CA, USA). .. Samples containing trace amounts of genomic DNA were treated with DNase I (Invitrogen™, Waltham, MA) and cleaned up using RNeasy MinElute Cleanup Kit (Qiagen, Hilden, Germany).

    Laser Capture Microdissection:

    Article Title: INTUSSUSCEPTIVE-LIKE ANGIOGENESIS IN HUMAN FETAL LUNG XENOGRAFTS: LINK WITH BRONCHOPULMONARY DYSPLASIA-ASSOCIATED MICROVASCULAR DYSANGIOGENESIS?
    Article Snippet: Paragraph title: Laser-capture microdissection, RNA extraction, microarray and qRT-PCR analysis of gene expression in xenografts ... RNA was further purified and concentrated using the RNEasy Minelute Cleanup kit (Qiagen, Valencia, CA) and then evaluated by the Agilent Bioanalyzer using an RNA 6000 Nano or Pico LabChip (Agilent Technologies, Santa Clara, CA).

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: Forty-eight hours post gene transfer, 400 ng/ml of puromycin was added, and the cells were expanded for 7 additional days followed by genomic DNA isolation. nrLAM PCR or LAM PCR with MseI or MluCI , , was performed, and deep sequencing data were generated with the Illumina MiSeq platform (San Diego, CA). .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    Random Hexamer Labeling:

    Article Title: Direct regulation of nucleosome density by the conserved AAA-ATPase Yta7
    Article Snippet: Total RNA was digested with DNase I (Roche) and purified using an RNeasy MinElute kit (Qiagen). .. RNA for the experiments illustrated in and and and was isolated directly from whole-cell extract using an RNeasy Mini kit (Qiagen) with on-column DNase I (Qiagen) digestion. cDNA was synthesized using a Super-Script III First-Strand Synthesis System (Invitrogen).

    Knock-Out:

    Article Title: Inferior olivary TMEM16B mediates cerebellar motor learning
    Article Snippet: For RT-PCR, retina, olfactory bulbs, hippocampi, and medulla oblongata were dissected from TMEM16B knockout mice and wild type controls (1.5 – 2 months old). .. Total RNA was extracted with Trizol® reagent (Life Technologies) and purified with RNeasy MinElute Kit (Qiagen) following manufacturers’ instructions.

    Produced:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    Concentration Assay:

    Article Title: Biomarkers from circulating neutrophil transcriptomes have potential to detect unruptured intracranial aneurysms
    Article Snippet: RNA was purified using the RNeasy MinElute Cleanup Kit (Qiagen, Venlo, Limburg, Netherlands) and suspended in RNase-free water. .. The purity and concentration of RNA in each sample were measured by absorbance at 260 nm on a NanoDrop 2000 spectrophotometer (Thermo Scientific, Waltham, MA), and 200–400 ng of RNA was sent to our university’s Next-Generation Sequencing and Expression Analysis Core facility for further quality control.

    Article Title: Direct and indirect parental exposure to endocrine disruptors and elevated temperature influences gene expression across generations in a euryhaline model fish
    Article Snippet: RNA was extracted using RNeasy Mini Kits (Qiagen, Hilden, Germany) Total RNA concentration and 260/280 ratios (acceptable values > 1.6) were quantified using a Nanodrop 1000 spectrophotometer (ThermoFisher Scientific, Waltham, MA). .. Samples containing trace amounts of genomic DNA were treated with DNase I (Invitrogen™, Waltham, MA) and cleaned up using RNeasy MinElute Cleanup Kit (Qiagen, Hilden, Germany).

    High Throughput Screening Assay:

    Article Title: Evaluation of TCR Gene Editing Achieved by TALENs, CRISPR/Cas9, and megaTAL Nucleases
    Article Snippet: The high-throughput insertion site analysis pipeline was utilized for vector trimming and genome alignment, and IS/CLIS identification was determined. .. Total RNA was isolated from MT-treated CD3-negative cells that were sorted to purity using the RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed with SuperScript Vilo (Thermo Fisher Scientific) and PCR amplified with primers: F:5′-GCCGAGGAGTCTTGTAAATGTAATGG-3′ and R:5′- TCACTTGTCAATTAATCCAGCTTCC-3′ at 98 °C × 2 minutes, 98 °C × 40 seconds, 58 °C × 40 seconds, 68 °C × 2 minutes and 30 seconds for 40 cycles.

    FACS:

    Article Title: Interactions of the Hdm2/p53 and Proteasome Pathways May Enhance the Antitumor Activity of Bortezomib
    Article Snippet: MM.1S-GFP/luc cells were cultured for 24 h in the presence or absence of the BMSC line HS-5 and then sorted by fluorescence activated cell sorting (FACS) cell sorter (BD FACSAria cell sorter, Becton Dickinson). .. RNA was extracted with TRIZOL-LS (Invitrogen), further treated with DNase I, and purified with the RNeasy MinElute Cleanup kit (Qiagen).

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    Qiagen rneasy minelute column
    Rneasy Minelute Column, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen rna affinity columns
    Functional and structural validation of newly discovered HIV-1 <t>RNA</t> motifs ( a ) Scheme for simultaneous deconvolution and structural analysis of a mixture of native sequence and U3 PK mutant genomes. (b) SHAPE profiles for the U3 PK pseudoknot bridging U3 and R. The experiment simultaneously probed a mixture of viruses with native sequence and mutant U3 PK <t>RNAs.</t> Secondary structure for the native sequence is shown as arcs below the y-axis intercept. Significant SHAPE reactivity differences are emphasized with yellow vertical lines (see Online Methods). (c) Direct growth competition and viral spread for U3 PK mutant and native sequence NL4-3 HIV-1 virions in Jurkat cells. Percentage of mutant in the initial inoculum is presented as a grey square at day 0. p24 levels correspond to the amount of HIV-1 capsid protein. ( d ) SHAPE profiles for the RT PK pseudoknot within the reverse transcriptase coding region. In this case, SHAPE data were obtained in separate experiments for each virus. ( e ) Viral spread and direct growth competition for RT PK mutant and native sequence NL4-3 HIV-1 virions in Jurkat cells. For the competition data, y-axes are shown on an expanded scale for clarity.
    Rna Affinity Columns, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Functional and structural validation of newly discovered HIV-1 RNA motifs ( a ) Scheme for simultaneous deconvolution and structural analysis of a mixture of native sequence and U3 PK mutant genomes. (b) SHAPE profiles for the U3 PK pseudoknot bridging U3 and R. The experiment simultaneously probed a mixture of viruses with native sequence and mutant U3 PK RNAs. Secondary structure for the native sequence is shown as arcs below the y-axis intercept. Significant SHAPE reactivity differences are emphasized with yellow vertical lines (see Online Methods). (c) Direct growth competition and viral spread for U3 PK mutant and native sequence NL4-3 HIV-1 virions in Jurkat cells. Percentage of mutant in the initial inoculum is presented as a grey square at day 0. p24 levels correspond to the amount of HIV-1 capsid protein. ( d ) SHAPE profiles for the RT PK pseudoknot within the reverse transcriptase coding region. In this case, SHAPE data were obtained in separate experiments for each virus. ( e ) Viral spread and direct growth competition for RT PK mutant and native sequence NL4-3 HIV-1 virions in Jurkat cells. For the competition data, y-axes are shown on an expanded scale for clarity.

    Journal: Nature methods

    Article Title: RNA motif discovery by SHAPE and mutational profiling (SHAPE-MaP)

    doi: 10.1038/nmeth.3029

    Figure Lengend Snippet: Functional and structural validation of newly discovered HIV-1 RNA motifs ( a ) Scheme for simultaneous deconvolution and structural analysis of a mixture of native sequence and U3 PK mutant genomes. (b) SHAPE profiles for the U3 PK pseudoknot bridging U3 and R. The experiment simultaneously probed a mixture of viruses with native sequence and mutant U3 PK RNAs. Secondary structure for the native sequence is shown as arcs below the y-axis intercept. Significant SHAPE reactivity differences are emphasized with yellow vertical lines (see Online Methods). (c) Direct growth competition and viral spread for U3 PK mutant and native sequence NL4-3 HIV-1 virions in Jurkat cells. Percentage of mutant in the initial inoculum is presented as a grey square at day 0. p24 levels correspond to the amount of HIV-1 capsid protein. ( d ) SHAPE profiles for the RT PK pseudoknot within the reverse transcriptase coding region. In this case, SHAPE data were obtained in separate experiments for each virus. ( e ) Viral spread and direct growth competition for RT PK mutant and native sequence NL4-3 HIV-1 virions in Jurkat cells. For the competition data, y-axes are shown on an expanded scale for clarity.

    Article Snippet: Following modification, RNAs were isolated using either RNA affinity columns (RNeasy MinElute; Qiagen) or G-50 spin columns (GE Healthcare).

    Techniques: Functional Assay, Sequencing, Mutagenesis