superscript ii reverse transcriptase  (Thermo Fisher)


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    Structured Review

    Thermo Fisher superscript ii reverse transcriptase
    Superscript Ii Reverse Transcriptase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 572 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript ii reverse transcriptase/product/Thermo Fisher
    Average 99 stars, based on 572 article reviews
    Price from $9.99 to $1999.99
    superscript ii reverse transcriptase - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Selective Regulation of N-Type Ca Channels by Different Combinations of G-Protein β/γ Subunits and RGS Proteins
    Article Snippet: Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ). β subunit cDNAs were amplified from the isolated mRNA by reverse transcription-PCR (thermal cycling: 98°C, 20 min; 56°C, 1 min; 72°C, 1.5 min; 35 cycles, preceded by a 3 min 98°C denaturing) with SuperScript II reverse transcriptase (Life Technologies, Rockville, MD) and oligo-dT oligonucleotides and then with specific primers and TakaRa LA Taq DNA polymerase (PanVera, Madison, WI). .. The resulting amplification products were first cloned into pCRII/TOPO vector (Invitrogen, San Diego, CA) for verification.

    Amplification:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: .. RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany). .. In vitro interaction assays.

    Article Title: Selective Regulation of N-Type Ca Channels by Different Combinations of G-Protein β/γ Subunits and RGS Proteins
    Article Snippet: .. Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ). β subunit cDNAs were amplified from the isolated mRNA by reverse transcription-PCR (thermal cycling: 98°C, 20 min; 56°C, 1 min; 72°C, 1.5 min; 35 cycles, preceded by a 3 min 98°C denaturing) with SuperScript II reverse transcriptase (Life Technologies, Rockville, MD) and oligo-dT oligonucleotides and then with specific primers and TakaRa LA Taq DNA polymerase (PanVera, Madison, WI). .. Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ).

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY). .. Conventional PCR amplification of cDNA was done in a 25 μL reaction volume by using DreamTaq Green DNA polymerase (Thermo Fisher Scientific).

    Article Title: An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1 [OPEN]
    Article Snippet: Three biological replicates of basal and apical root segments at each time point were subjected to RNA extraction using an RNeasy plant mini kit (Qiagen) with on-column DNase treatment according to the manufacturer’s instructions. cDNA was prepared using random hexamer primers and SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer’s protocol. .. RNA amplification, labeling, and hybridization to 60K Agilent custom-made gene expression microarrays (design available at EMBL-EBI ArrayExpress, accession no. A-MTAB-530) were conducted as described ( ).

    Synthesized:

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: .. RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.). .. QRT-PCR analysis was performed on a TaqMan® ABI Prism 7000 Sequence Detection System (Applied Biosystems, Foster City, CA).

    Quantitative RT-PCR:

    Article Title: The histone demethylase LSD1 regulates B cell proliferation and plasmablast differentiation
    Article Snippet: Paragraph title: qRT-PCR ... Total RNA was reverse transcribed with SuperScript II reverse transcriptase (Invitrogen 18064014). cDNA was diluted five-fold and qPCR was performed with a CFX96 instrument (BioRad) using SYBR Green incorporation.

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: Paragraph title: TaqMan® quantitative real-time PCR (QRT-PCR) ... RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.).

    Article Title: Loss of Tbx3 in murine neural crest reduces enteric glia and causes cleft palate, but does not influence heart development or bowel transit
    Article Snippet: .. Total RNA was isolated from FACS sorted bowel cells or microdissected palatal shelves at E13.5 using RNeasy Mini Kit (Qiagen #74104) or RNEasy Micro Kit Qiagen #74004 and reverse-transcribed using Superscript II Reverse Transcriptase for cDNA synthesis (Invitrogen #18064014). qRT-PCR was performed on at least three biological replicates with three technical replicates per run using SsoFast™ EvaGreen® Supermix with Low ROX (Bio-Rad #1725211) and Bio-Rad Cycler CFX96. .. Photographs of whole fetuses, skeletal preparations, and hearts were acquired on an Olympus SZ40 stereomicroscope.

    Article Title: Rab34 small GTPase is required for Hedgehog signaling and an early step of ciliary vesicle formation in mouse
    Article Snippet: Of the isolated total RNAs, 5μg of each were used to synthesize first stranded cDNA using SuperScript II reverse transcriptase according to the manufacturer's instruction (Thermo Fisher Scientific). .. Reverse transcription quantitative PCR (RT-qPCR) was performed using a qPCR kit (MasterMix-R, Applied Biological Materials, Inc., Canada) and primers described previously ( ).

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: Gene expression at the mRNA level was evaluated by conventional RT-PCR, quantitative RT-PCR (RT-qPCR), and RNA sequencing (RNA-seq) analyses. .. Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY).

    SYBR Green Assay:

    Article Title: The histone demethylase LSD1 regulates B cell proliferation and plasmablast differentiation
    Article Snippet: .. Total RNA was reverse transcribed with SuperScript II reverse transcriptase (Invitrogen 18064014). cDNA was diluted five-fold and qPCR was performed with a CFX96 instrument (BioRad) using SYBR Green incorporation. .. Gene expression was calculated relative to ERCC transcript #130.

    Article Title: Genetic Analysis of Myc and Telomerase Interactions In Vivo
    Article Snippet: Briefly, reverse transcription was conducted with 1 μg of total RNA, random hexamers as primers, and Superscript II reverse transcriptase (Invitrogen). .. Real-time PCR was done with two dilutions in duplicates of cDNA on an ABI Prism 7700 Sequence Detector (Applied Biosystems), with SYBR Green (Applied Biosystems), where each reaction contained 1× SYBR-Green mix, 3 mM MgCl2 , 0.4 μM each primer, 0.5 mM deoxynucleoside triphosphates, and 5 μl of the sample.

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY). .. RT-qPCR amplification of cDNAs was carried out in a 20-μL reaction mixture containing Applied Biosystems Power SYBR Green PCR Master Mix (Thermo Fisher Scientific).

    Microarray:

    Article Title: An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1 [OPEN]
    Article Snippet: Three biological replicates of basal and apical root segments at each time point were subjected to RNA extraction using an RNeasy plant mini kit (Qiagen) with on-column DNase treatment according to the manufacturer’s instructions. cDNA was prepared using random hexamer primers and SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer’s protocol. .. Analysis of microarray data were performed with the R package limma ( ).

    Incubation:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: Cells were incubated with or without 100 μM DRB for 1 h and immunoprecipitations and western blotting were carried out as described previously ( ). .. RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany).

    Article Title: Rab34 small GTPase is required for Hedgehog signaling and an early step of ciliary vesicle formation in mouse
    Article Snippet: Confluent WT and Rab34-mutant pMEFs in 60†mm plates were incubated with growth medium containing SAG (200†nM; Cayman Chemical) or vehicle control overnight. .. Of the isolated total RNAs, 5μg of each were used to synthesize first stranded cDNA using SuperScript II reverse transcriptase according to the manufacturer's instruction (Thermo Fisher Scientific).

    Expressing:

    Article Title: The histone demethylase LSD1 regulates B cell proliferation and plasmablast differentiation
    Article Snippet: Total RNA was reverse transcribed with SuperScript II reverse transcriptase (Invitrogen 18064014). cDNA was diluted five-fold and qPCR was performed with a CFX96 instrument (BioRad) using SYBR Green incorporation. .. Gene expression was calculated relative to ERCC transcript #130.

    Article Title: Selective Regulation of N-Type Ca Channels by Different Combinations of G-Protein β/γ Subunits and RGS Proteins
    Article Snippet: Expression constructs. .. Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ). β subunit cDNAs were amplified from the isolated mRNA by reverse transcription-PCR (thermal cycling: 98°C, 20 min; 56°C, 1 min; 72°C, 1.5 min; 35 cycles, preceded by a 3 min 98°C denaturing) with SuperScript II reverse transcriptase (Life Technologies, Rockville, MD) and oligo-dT oligonucleotides and then with specific primers and TakaRa LA Taq DNA polymerase (PanVera, Madison, WI).

    Article Title: Genetic Analysis of Myc and Telomerase Interactions In Vivo
    Article Snippet: Briefly, reverse transcription was conducted with 1 μg of total RNA, random hexamers as primers, and Superscript II reverse transcriptase (Invitrogen). .. For each mRNA sample, mTert expression was corrected by the actin mRNA content in each sample.

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: Paragraph title: Detection of gene expression at mRNA level ... Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY).

    Article Title: An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1 [OPEN]
    Article Snippet: Three biological replicates of basal and apical root segments at each time point were subjected to RNA extraction using an RNeasy plant mini kit (Qiagen) with on-column DNase treatment according to the manufacturer’s instructions. cDNA was prepared using random hexamer primers and SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer’s protocol. .. RNA amplification, labeling, and hybridization to 60K Agilent custom-made gene expression microarrays (design available at EMBL-EBI ArrayExpress, accession no. A-MTAB-530) were conducted as described ( ).

    Western Blot:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: Cells were incubated with or without 100 μM DRB for 1 h and immunoprecipitations and western blotting were carried out as described previously ( ). .. RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany).

    Translocation Assay:

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: BMP2- or BMP4 treatment of MEMM cells was performed as described below under “BMP treatment of MEMM cells and determination of nuclear translocation of BR-Smad proteins.” The quality and quantity of extracted total RNAs were assessed by formaldehyde agarose gel electrophoresis and spectrophotometric UV absorbance at 260/280 nm, respectively. .. RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.).

    Hybridization:

    Article Title: An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1 [OPEN]
    Article Snippet: Three biological replicates of basal and apical root segments at each time point were subjected to RNA extraction using an RNeasy plant mini kit (Qiagen) with on-column DNase treatment according to the manufacturer’s instructions. cDNA was prepared using random hexamer primers and SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer’s protocol. .. RNA amplification, labeling, and hybridization to 60K Agilent custom-made gene expression microarrays (design available at EMBL-EBI ArrayExpress, accession no. A-MTAB-530) were conducted as described ( ).

    Two Tailed Test:

    Article Title: The histone demethylase LSD1 regulates B cell proliferation and plasmablast differentiation
    Article Snippet: Total RNA was reverse transcribed with SuperScript II reverse transcriptase (Invitrogen 18064014). cDNA was diluted five-fold and qPCR was performed with a CFX96 instrument (BioRad) using SYBR Green incorporation. .. A Student’s two-tailed t-test was used to determine statistical significance.

    Polymerase Chain Reaction:

    Article Title: Histone H3 tail clipping regulates gene expression
    Article Snippet: Paragraph title: Quantitative reverse-transcription PCR ... First strand cDNA synthesis was achieved using SuperScript II reverse transcriptase (cat.18080, Invitrogen) with a primer cocktail, containing 50 uM oligo(dT) (Ambion) and 50 ng random hexamers (Invitrogen), as described in the manufacturer’s instructions.

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY). .. Conventional PCR amplification of cDNA was done in a 25 μL reaction volume by using DreamTaq Green DNA polymerase (Thermo Fisher Scientific).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: .. RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany). .. In vitro interaction assays.

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: Gene expression at the mRNA level was evaluated by conventional RT-PCR, quantitative RT-PCR (RT-qPCR), and RNA sequencing (RNA-seq) analyses. .. Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY).

    Affinity Purification:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: Polyclonal antibodies against LARP7 were raised by immunizing rabbits with bacterially expressed full-length 6 × His-LARP7 and affinity purified. .. RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany).

    Binding Assay:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: In vitro -transcribed 32 P-labelled wild-type or mutant 7SK RNAs were incubated with 250 ng of either recombinant full-length LARP7 or a fragment thereof (amino acids 1–197) for 20 min at 20°C in binding buffer (10 mM HEPES, pH 7.5, 10 mM KCl, 2 mM MgCl2 , 1 mM dithiothreitol, 5% glycerol and 0.1 μg transfer RNA). .. RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany).

    RNA Sequencing Assay:

    Article Title: The histone demethylase LSD1 regulates B cell proliferation and plasmablast differentiation
    Article Snippet: Per sample, 50,000 cells were sorted via FACS into 600 μl RLT-βME and processed as with RNA-seq. .. Total RNA was reverse transcribed with SuperScript II reverse transcriptase (Invitrogen 18064014). cDNA was diluted five-fold and qPCR was performed with a CFX96 instrument (BioRad) using SYBR Green incorporation.

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: Gene expression at the mRNA level was evaluated by conventional RT-PCR, quantitative RT-PCR (RT-qPCR), and RNA sequencing (RNA-seq) analyses. .. Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY).

    Fluorescence:

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.). .. Primers and their corresponding fluorescence probes (Assays-on-Demand) were purchased from Applied Biosystems.

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY). .. Amplification and fluorescence detection of RT-qPCR were carried out on Applied Biosystems QuantStudio Flex 7 Real Time PCR System (Thermo Fisher Scientific).

    Mutagenesis:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: In vitro -transcribed 32 P-labelled wild-type or mutant 7SK RNAs were incubated with 250 ng of either recombinant full-length LARP7 or a fragment thereof (amino acids 1–197) for 20 min at 20°C in binding buffer (10 mM HEPES, pH 7.5, 10 mM KCl, 2 mM MgCl2 , 1 mM dithiothreitol, 5% glycerol and 0.1 μg transfer RNA). .. RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany).

    Isolation:

    Article Title: Selective Regulation of N-Type Ca Channels by Different Combinations of G-Protein β/γ Subunits and RGS Proteins
    Article Snippet: .. Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ). β subunit cDNAs were amplified from the isolated mRNA by reverse transcription-PCR (thermal cycling: 98°C, 20 min; 56°C, 1 min; 72°C, 1.5 min; 35 cycles, preceded by a 3 min 98°C denaturing) with SuperScript II reverse transcriptase (Life Technologies, Rockville, MD) and oligo-dT oligonucleotides and then with specific primers and TakaRa LA Taq DNA polymerase (PanVera, Madison, WI). .. Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ).

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: Total RNA from BMP2- or BMP4- or vehicle-treated MEMM cells was isolated as noted above. .. RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.).

    Article Title: Loss of Tbx3 in murine neural crest reduces enteric glia and causes cleft palate, but does not influence heart development or bowel transit
    Article Snippet: .. Total RNA was isolated from FACS sorted bowel cells or microdissected palatal shelves at E13.5 using RNeasy Mini Kit (Qiagen #74104) or RNEasy Micro Kit Qiagen #74004 and reverse-transcribed using Superscript II Reverse Transcriptase for cDNA synthesis (Invitrogen #18064014). qRT-PCR was performed on at least three biological replicates with three technical replicates per run using SsoFast™ EvaGreen® Supermix with Low ROX (Bio-Rad #1725211) and Bio-Rad Cycler CFX96. .. Photographs of whole fetuses, skeletal preparations, and hearts were acquired on an Olympus SZ40 stereomicroscope.

    Article Title: Hypoxia-induced Deoxycytidine Kinase Contributes to Epithelial Proliferation in Pulmonary Fibrosis
    Article Snippet: Total RNA was isolated from MLE12 cells or frozen lungtissue using TRIzol (Life Technologies). .. RNA was DNase treated andreverse-transcribed using Superscript II reverse transcriptase (Life Technologies).Mouse β-actin or 18S ribosomal RNA was used as internal controls.

    Article Title: Rab34 small GTPase is required for Hedgehog signaling and an early step of ciliary vesicle formation in mouse
    Article Snippet: .. Of the isolated total RNAs, 5μg of each were used to synthesize first stranded cDNA using SuperScript II reverse transcriptase according to the manufacturer's instruction (Thermo Fisher Scientific). .. Reverse transcription quantitative PCR (RT-qPCR) was performed using a qPCR kit (MasterMix-R, Applied Biological Materials, Inc., Canada) and primers described previously ( ).

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: .. Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY). .. Conventional PCR amplification of cDNA was done in a 25 μL reaction volume by using DreamTaq Green DNA polymerase (Thermo Fisher Scientific).

    Article Title: Signals in Stem Cell Differentiation on Fluorapatite-Modified Scaffolds
    Article Snippet: Paragraph title: RNA Isolation and Reverse Transcription ... For converting the total RNA into cDNA for RNA array samples, the RT2 First Stand cDNA Synthesis and the SuperScript II Reverse Transcriptase (Invitrogen) kits were used following manufacturer’s protocols.

    Labeling:

    Article Title: An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1 [OPEN]
    Article Snippet: Three biological replicates of basal and apical root segments at each time point were subjected to RNA extraction using an RNeasy plant mini kit (Qiagen) with on-column DNase treatment according to the manufacturer’s instructions. cDNA was prepared using random hexamer primers and SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer’s protocol. .. RNA amplification, labeling, and hybridization to 60K Agilent custom-made gene expression microarrays (design available at EMBL-EBI ArrayExpress, accession no. A-MTAB-530) were conducted as described ( ).

    Purification:

    Article Title: Selective Regulation of N-Type Ca Channels by Different Combinations of G-Protein β/γ Subunits and RGS Proteins
    Article Snippet: .. Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ). β subunit cDNAs were amplified from the isolated mRNA by reverse transcription-PCR (thermal cycling: 98°C, 20 min; 56°C, 1 min; 72°C, 1.5 min; 35 cycles, preceded by a 3 min 98°C denaturing) with SuperScript II reverse transcriptase (Life Technologies, Rockville, MD) and oligo-dT oligonucleotides and then with specific primers and TakaRa LA Taq DNA polymerase (PanVera, Madison, WI). .. Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ).

    Sequencing:

    Article Title: Selective Regulation of N-Type Ca Channels by Different Combinations of G-Protein β/γ Subunits and RGS Proteins
    Article Snippet: Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ). β subunit cDNAs were amplified from the isolated mRNA by reverse transcription-PCR (thermal cycling: 98°C, 20 min; 56°C, 1 min; 72°C, 1.5 min; 35 cycles, preceded by a 3 min 98°C denaturing) with SuperScript II reverse transcriptase (Life Technologies, Rockville, MD) and oligo-dT oligonucleotides and then with specific primers and TakaRa LA Taq DNA polymerase (PanVera, Madison, WI). .. All constructs contained an expression-optimized Kozak sequence (GCCACC) before the 5′-ATG start codon.

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.). .. QRT-PCR analysis was performed on a TaqMan® ABI Prism 7000 Sequence Detection System (Applied Biosystems, Foster City, CA).

    Article Title: Genetic Analysis of Myc and Telomerase Interactions In Vivo
    Article Snippet: Briefly, reverse transcription was conducted with 1 μg of total RNA, random hexamers as primers, and Superscript II reverse transcriptase (Invitrogen). .. Real-time PCR was done with two dilutions in duplicates of cDNA on an ABI Prism 7700 Sequence Detector (Applied Biosystems), with SYBR Green (Applied Biosystems), where each reaction contained 1× SYBR-Green mix, 3 mM MgCl2 , 0.4 μM each primer, 0.5 mM deoxynucleoside triphosphates, and 5 μl of the sample.

    Construct:

    Article Title: Selective Regulation of N-Type Ca Channels by Different Combinations of G-Protein β/γ Subunits and RGS Proteins
    Article Snippet: Expression constructs. .. Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ). β subunit cDNAs were amplified from the isolated mRNA by reverse transcription-PCR (thermal cycling: 98°C, 20 min; 56°C, 1 min; 72°C, 1.5 min; 35 cycles, preceded by a 3 min 98°C denaturing) with SuperScript II reverse transcriptase (Life Technologies, Rockville, MD) and oligo-dT oligonucleotides and then with specific primers and TakaRa LA Taq DNA polymerase (PanVera, Madison, WI).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Histone H3 tail clipping regulates gene expression
    Article Snippet: .. First strand cDNA synthesis was achieved using SuperScript II reverse transcriptase (cat.18080, Invitrogen) with a primer cocktail, containing 50 uM oligo(dT) (Ambion) and 50 ng random hexamers (Invitrogen), as described in the manufacturer’s instructions. .. The cDNA samples were then used as templates for real-time PCR (see above).

    Mouse Assay:

    Article Title: Genetic Analysis of Myc and Telomerase Interactions In Vivo
    Article Snippet: Three or four 8-week-old mice of the indicated genotypes were treated by daily topical application of 4-OHT or EtOH to the upper dorsal skin. .. Briefly, reverse transcription was conducted with 1 μg of total RNA, random hexamers as primers, and Superscript II reverse transcriptase (Invitrogen).

    Plasmid Preparation:

    Article Title: Selective Regulation of N-Type Ca Channels by Different Combinations of G-Protein β/γ Subunits and RGS Proteins
    Article Snippet: Human Gβ1 , Gβ2 , Gβ3 , and Gβ5 ( ) were obtained from human embryonic kidney (HEK) cell line mRNA (Quick Prep Micro mRNA purification kit; Amersham Pharmacia Biotech, Piscataway, NJ). β subunit cDNAs were amplified from the isolated mRNA by reverse transcription-PCR (thermal cycling: 98°C, 20 min; 56°C, 1 min; 72°C, 1.5 min; 35 cycles, preceded by a 3 min 98°C denaturing) with SuperScript II reverse transcriptase (Life Technologies, Rockville, MD) and oligo-dT oligonucleotides and then with specific primers and TakaRa LA Taq DNA polymerase (PanVera, Madison, WI). .. The resulting amplification products were first cloned into pCRII/TOPO vector (Invitrogen, San Diego, CA) for verification.

    Real-time Polymerase Chain Reaction:

    Article Title: Histone H3 tail clipping regulates gene expression
    Article Snippet: First strand cDNA synthesis was achieved using SuperScript II reverse transcriptase (cat.18080, Invitrogen) with a primer cocktail, containing 50 uM oligo(dT) (Ambion) and 50 ng random hexamers (Invitrogen), as described in the manufacturer’s instructions. .. The cDNA samples were then used as templates for real-time PCR (see above).

    Article Title: The histone demethylase LSD1 regulates B cell proliferation and plasmablast differentiation
    Article Snippet: .. Total RNA was reverse transcribed with SuperScript II reverse transcriptase (Invitrogen 18064014). cDNA was diluted five-fold and qPCR was performed with a CFX96 instrument (BioRad) using SYBR Green incorporation. .. Gene expression was calculated relative to ERCC transcript #130.

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: Paragraph title: TaqMan® quantitative real-time PCR (QRT-PCR) ... RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.).

    Article Title: Genetic Analysis of Myc and Telomerase Interactions In Vivo
    Article Snippet: Paragraph title: Real-time quantitative PCR for mTert mRNA detection. ... Briefly, reverse transcription was conducted with 1 μg of total RNA, random hexamers as primers, and Superscript II reverse transcriptase (Invitrogen).

    Article Title: Hypoxia-induced Deoxycytidine Kinase Contributes to Epithelial Proliferation in Pulmonary Fibrosis
    Article Snippet: Paragraph title: Quantitative Real-Time Polymerase Chain Reaction ... RNA was DNase treated andreverse-transcribed using Superscript II reverse transcriptase (Life Technologies).Mouse β-actin or 18S ribosomal RNA was used as internal controls.

    Article Title: Rab34 small GTPase is required for Hedgehog signaling and an early step of ciliary vesicle formation in mouse
    Article Snippet: Paragraph title: Reverse transcription quantitative PCR ... Of the isolated total RNAs, 5μg of each were used to synthesize first stranded cDNA using SuperScript II reverse transcriptase according to the manufacturer's instruction (Thermo Fisher Scientific).

    Article Title: ESR2 Is Essential for Gonadotropin-Induced Kiss1 Expression in Granulosa Cells
    Article Snippet: Total RNA was isolated from the granulosa cells with TRI Reagent (Sigma-Aldrich) and cDNAs were prepared from 2 µg of total RNA with oligo random primers and Superscript II reverse transcriptase (Thermo Fisher Scientific, Grand Island, NY). .. Amplification and fluorescence detection of RT-qPCR were carried out on Applied Biosystems QuantStudio Flex 7 Real Time PCR System (Thermo Fisher Scientific).

    Article Title: An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1 [OPEN]
    Article Snippet: Paragraph title: Transcriptome Profiling and Quantitative PCR ... Three biological replicates of basal and apical root segments at each time point were subjected to RNA extraction using an RNeasy plant mini kit (Qiagen) with on-column DNase treatment according to the manufacturer’s instructions. cDNA was prepared using random hexamer primers and SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer’s protocol.

    RNA Extraction:

    Article Title: An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1 [OPEN]
    Article Snippet: .. Three biological replicates of basal and apical root segments at each time point were subjected to RNA extraction using an RNeasy plant mini kit (Qiagen) with on-column DNase treatment according to the manufacturer’s instructions. cDNA was prepared using random hexamer primers and SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer’s protocol. .. RNA amplification, labeling, and hybridization to 60K Agilent custom-made gene expression microarrays (design available at EMBL-EBI ArrayExpress, accession no. A-MTAB-530) were conducted as described ( ).

    Recombinant:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: In vitro -transcribed 32 P-labelled wild-type or mutant 7SK RNAs were incubated with 250 ng of either recombinant full-length LARP7 or a fragment thereof (amino acids 1–197) for 20 min at 20°C in binding buffer (10 mM HEPES, pH 7.5, 10 mM KCl, 2 mM MgCl2 , 1 mM dithiothreitol, 5% glycerol and 0.1 μg transfer RNA). .. RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany).

    Agarose Gel Electrophoresis:

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: BMP2- or BMP4 treatment of MEMM cells was performed as described below under “BMP treatment of MEMM cells and determination of nuclear translocation of BR-Smad proteins.” The quality and quantity of extracted total RNAs were assessed by formaldehyde agarose gel electrophoresis and spectrophotometric UV absorbance at 260/280 nm, respectively. .. RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.).

    In Vitro:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: In vitro -transcribed 32 P-labelled wild-type or mutant 7SK RNAs were incubated with 250 ng of either recombinant full-length LARP7 or a fragment thereof (amino acids 1–197) for 20 min at 20°C in binding buffer (10 mM HEPES, pH 7.5, 10 mM KCl, 2 mM MgCl2 , 1 mM dithiothreitol, 5% glycerol and 0.1 μg transfer RNA). .. RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany).

    Random Hexamer Labeling:

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: .. RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.). .. QRT-PCR analysis was performed on a TaqMan® ABI Prism 7000 Sequence Detection System (Applied Biosystems, Foster City, CA).

    Article Title: An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1An Age-Dependent Sequence of Physiological Processes Defines Developmental Root Senescence 1 [OPEN]
    Article Snippet: .. Three biological replicates of basal and apical root segments at each time point were subjected to RNA extraction using an RNeasy plant mini kit (Qiagen) with on-column DNase treatment according to the manufacturer’s instructions. cDNA was prepared using random hexamer primers and SuperScript II reverse transcriptase (Life Technologies) according to the manufacturer’s protocol. .. RNA amplification, labeling, and hybridization to 60K Agilent custom-made gene expression microarrays (design available at EMBL-EBI ArrayExpress, accession no. A-MTAB-530) were conducted as described ( ).

    Produced:

    Article Title: The La-related protein LARP7 is a component of the 7SK ribonucleoprotein and affects transcription of cellular and viral polymerase II genes
    Article Snippet: RNA was phenol extracted and amplified by RT–PCR with superscript II reverse transcriptase (Invitrogen, Karlsrune, Germany). .. 35 S-labelled proteins were produced as reported previously ( ).

    Concentration Assay:

    Article Title: BMP Signaling Dynamics in Embryonic Orofacial Tissue
    Article Snippet: RNA was treated with DNase I in the presence of RNaseOUT (Invitrogen Life Technologies, Inc.) to remove contaminating DNA before cDNA synthesis. cDNA was synthesized from total RNA using random hexamer primers and Superscript II reverse transcriptase (Invitrogen Life Technologies, Inc.). .. For each gene analyzed, both forward and reverse primers were used at a concentration of 900 nM and the final fluorescent probe concentration was 200 nM.

    CTG Assay:

    Article Title: Genetic Analysis of Myc and Telomerase Interactions In Vivo
    Article Snippet: Briefly, reverse transcription was conducted with 1 μg of total RNA, random hexamers as primers, and Superscript II reverse transcriptase (Invitrogen). .. The PCRs employed a set of primers specific for the mTert gene (TERT-F, 5′ GGA TTG CCA CTG GCT CCG 3′; TERT-R, 5′ TGC CTG ACC TCC TCT TGT GAC 3′) and a set specific for actin (ACTIN-F, 5′ GGC ACC ACA CCT TCT ACA ATG 3′, ACTIN-R, 5′ GTG GTG GTG AAG CTG TAG 3′).

    FACS:

    Article Title: The histone demethylase LSD1 regulates B cell proliferation and plasmablast differentiation
    Article Snippet: Per sample, 50,000 cells were sorted via FACS into 600 μl RLT-βME and processed as with RNA-seq. .. Total RNA was reverse transcribed with SuperScript II reverse transcriptase (Invitrogen 18064014). cDNA was diluted five-fold and qPCR was performed with a CFX96 instrument (BioRad) using SYBR Green incorporation.

    Article Title: Loss of Tbx3 in murine neural crest reduces enteric glia and causes cleft palate, but does not influence heart development or bowel transit
    Article Snippet: .. Total RNA was isolated from FACS sorted bowel cells or microdissected palatal shelves at E13.5 using RNeasy Mini Kit (Qiagen #74104) or RNEasy Micro Kit Qiagen #74004 and reverse-transcribed using Superscript II Reverse Transcriptase for cDNA synthesis (Invitrogen #18064014). qRT-PCR was performed on at least three biological replicates with three technical replicates per run using SsoFast™ EvaGreen® Supermix with Low ROX (Bio-Rad #1725211) and Bio-Rad Cycler CFX96. .. Photographs of whole fetuses, skeletal preparations, and hearts were acquired on an Olympus SZ40 stereomicroscope.

    T-Test:

    Article Title: The histone demethylase LSD1 regulates B cell proliferation and plasmablast differentiation
    Article Snippet: Total RNA was reverse transcribed with SuperScript II reverse transcriptase (Invitrogen 18064014). cDNA was diluted five-fold and qPCR was performed with a CFX96 instrument (BioRad) using SYBR Green incorporation. .. A Student’s two-tailed t-test was used to determine statistical significance.

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  • 99
    Thermo Fisher dnase i
    Influence of Eap on DNA stability. (A) DNase activities of different Eap (3 μg/ml) preparations. Eap samples were purified from strain Newman (New), its nuc1 nuc2 derivative (M0746N1), or as a recombinant protein from E. coli (rMu50). DNase activity was quantified by ELISA (given in optical density (OD) readings at 450 nm). PCR grade water and <t>DNase</t> I (12.5 mKuU) served as negative and positive controls, respectively. OD 450 nm readings were normalized in relation to the values obtained for the negative controls, which were set to 100%. Data represent the mean ± SD of four to six independent experiments. ** P
    Dnase I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnase i/product/Thermo Fisher
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    dnase i - by Bioz Stars, 2020-04
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    99
    Thermo Fisher superscript ii reverse transcriptase
    Influence of Eap on DNA stability. (A) DNase activities of different Eap (3 μg/ml) preparations. Eap samples were purified from strain Newman (New), its nuc1 nuc2 derivative (M0746N1), or as a recombinant protein from E. coli (rMu50). DNase activity was quantified by ELISA (given in optical density (OD) readings at 450 nm). PCR grade water and <t>DNase</t> I (12.5 mKuU) served as negative and positive controls, respectively. OD 450 nm readings were normalized in relation to the values obtained for the negative controls, which were set to 100%. Data represent the mean ± SD of four to six independent experiments. ** P
    Superscript Ii Reverse Transcriptase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 572 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript ii reverse transcriptase/product/Thermo Fisher
    Average 99 stars, based on 572 article reviews
    Price from $9.99 to $1999.99
    superscript ii reverse transcriptase - by Bioz Stars, 2020-04
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    Influence of Eap on DNA stability. (A) DNase activities of different Eap (3 μg/ml) preparations. Eap samples were purified from strain Newman (New), its nuc1 nuc2 derivative (M0746N1), or as a recombinant protein from E. coli (rMu50). DNase activity was quantified by ELISA (given in optical density (OD) readings at 450 nm). PCR grade water and DNase I (12.5 mKuU) served as negative and positive controls, respectively. OD 450 nm readings were normalized in relation to the values obtained for the negative controls, which were set to 100%. Data represent the mean ± SD of four to six independent experiments. ** P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: The Staphylococcus aureus Extracellular Adherence Protein Eap Is a DNA Binding Protein Capable of Blocking Neutrophil Extracellular Trap Formation

    doi: 10.3389/fcimb.2018.00235

    Figure Lengend Snippet: Influence of Eap on DNA stability. (A) DNase activities of different Eap (3 μg/ml) preparations. Eap samples were purified from strain Newman (New), its nuc1 nuc2 derivative (M0746N1), or as a recombinant protein from E. coli (rMu50). DNase activity was quantified by ELISA (given in optical density (OD) readings at 450 nm). PCR grade water and DNase I (12.5 mKuU) served as negative and positive controls, respectively. OD 450 nm readings were normalized in relation to the values obtained for the negative controls, which were set to 100%. Data represent the mean ± SD of four to six independent experiments. ** P

    Article Snippet: Subsequently, cells were challenged with increasing concentrations of Eap (0, 1, 2.5, and 10 μg/ml) or DNase I (10 U/ml; Thermo Fisher), and incubated for 3 h at 37°C in presence of 5% CO2 .

    Techniques: Purification, Recombinant, Activity Assay, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction

    TLR9 signalling pathway, miR‐7 and Smad2 expressions in NET‐treated LFs. LFs were divided into control group, PMA group and PMA+DNase I group. A, Western blot assay showed that TLR9 protein level and phosphorylation levels of its downstream molecules p38, AKT and p65 were up‐regulated in PMA group, whereas PMA+DNase I decreased these expressions. B, miR‐7 expression level was down‐regulated in PMA group, whereas PMA+DNase I increased miR‐7 expression. C, Smad2 protein level was up‐regulated in PMA group, whereas PMA+DNase I inhibited Smad2 expression. ** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Neutrophil extracellular traps activate lung fibroblast to induce polymyositis‐related interstitial lung diseases via TLR9‐miR‐7‐Smad2 pathway, et al. Neutrophil extracellular traps activate lung fibroblast to induce polymyositis‐related interstitial lung diseases via TLR9‐miR‐7‐Smad2 pathway

    doi: 10.1111/jcmm.14858

    Figure Lengend Snippet: TLR9 signalling pathway, miR‐7 and Smad2 expressions in NET‐treated LFs. LFs were divided into control group, PMA group and PMA+DNase I group. A, Western blot assay showed that TLR9 protein level and phosphorylation levels of its downstream molecules p38, AKT and p65 were up‐regulated in PMA group, whereas PMA+DNase I decreased these expressions. B, miR‐7 expression level was down‐regulated in PMA group, whereas PMA+DNase I increased miR‐7 expression. C, Smad2 protein level was up‐regulated in PMA group, whereas PMA+DNase I inhibited Smad2 expression. ** P

    Article Snippet: In PMA+DNase I group, the LFs were cultivated by PMA‐stimulated NETs pre‐treated with DNase I (10 U/mL, Thermo Scientific) in 37°C for 30 minutes.

    Techniques: Western Blot, Expressing

    NETs accelerated the proliferation of LF and their differentiation into MF. LFs were divided into control group, PMA group and PMA+DNase I group. A, SYTOX Green nucleic acid stain showed that the nuclear membrane was fractured and DNA was released in PMA‐treated NET. B, qRT‐PCR assay detected mRNA levels of ACTA2 (α‐SMA gene), CCN2 (connective tissue growth factor) and ADAM12 (ADAM metallopeptidase domain 12) in control, PMA and PMA+DNase I groups. C, Western blot assay detected protein levels of α‐SMA and CCN2 in control, PMA and PMA+DNase I groups. D, PMA‐stimulated NET promoted the formation of collagen in LF, whereas the formation of collagen was reduced by DNase I. E‐F, The proliferation of LF was detected by MTT assay. G, The proliferation of LF was also detected by EdU experiment. H, The apoptosis of LF was detected by flow cytometry analysis. ** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Neutrophil extracellular traps activate lung fibroblast to induce polymyositis‐related interstitial lung diseases via TLR9‐miR‐7‐Smad2 pathway, et al. Neutrophil extracellular traps activate lung fibroblast to induce polymyositis‐related interstitial lung diseases via TLR9‐miR‐7‐Smad2 pathway

    doi: 10.1111/jcmm.14858

    Figure Lengend Snippet: NETs accelerated the proliferation of LF and their differentiation into MF. LFs were divided into control group, PMA group and PMA+DNase I group. A, SYTOX Green nucleic acid stain showed that the nuclear membrane was fractured and DNA was released in PMA‐treated NET. B, qRT‐PCR assay detected mRNA levels of ACTA2 (α‐SMA gene), CCN2 (connective tissue growth factor) and ADAM12 (ADAM metallopeptidase domain 12) in control, PMA and PMA+DNase I groups. C, Western blot assay detected protein levels of α‐SMA and CCN2 in control, PMA and PMA+DNase I groups. D, PMA‐stimulated NET promoted the formation of collagen in LF, whereas the formation of collagen was reduced by DNase I. E‐F, The proliferation of LF was detected by MTT assay. G, The proliferation of LF was also detected by EdU experiment. H, The apoptosis of LF was detected by flow cytometry analysis. ** P

    Article Snippet: In PMA+DNase I group, the LFs were cultivated by PMA‐stimulated NETs pre‐treated with DNase I (10 U/mL, Thermo Scientific) in 37°C for 30 minutes.

    Techniques: Staining, Quantitative RT-PCR, Western Blot, MTT Assay, Flow Cytometry

    Influence of Eap on DNA stability. (A) DNase activities of different Eap (3 μg/ml) preparations. Eap samples were purified from strain Newman (New), its nuc1 nuc2 derivative (M0746N1), or as a recombinant protein from E. coli (rMu50). DNase activity was quantified by ELISA (given in optical density (OD) readings at 450 nm). PCR grade water and DNase I (12.5 mKuU) served as negative and positive controls, respectively. OD 450 nm readings were normalized in relation to the values obtained for the negative controls, which were set to 100%. Data represent the mean ± SD of four to six independent experiments. ** P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: The Staphylococcus aureus Extracellular Adherence Protein Eap Is a DNA Binding Protein Capable of Blocking Neutrophil Extracellular Trap Formation

    doi: 10.3389/fcimb.2018.00235

    Figure Lengend Snippet: Influence of Eap on DNA stability. (A) DNase activities of different Eap (3 μg/ml) preparations. Eap samples were purified from strain Newman (New), its nuc1 nuc2 derivative (M0746N1), or as a recombinant protein from E. coli (rMu50). DNase activity was quantified by ELISA (given in optical density (OD) readings at 450 nm). PCR grade water and DNase I (12.5 mKuU) served as negative and positive controls, respectively. OD 450 nm readings were normalized in relation to the values obtained for the negative controls, which were set to 100%. Data represent the mean ± SD of four to six independent experiments. ** P

    Article Snippet: Subsequently, cells were challenged with increasing concentrations of Eap (0, 1, 2.5, and 10 μg/ml) or DNase I (10 U/ml; Thermo Fisher), and incubated for 3 h at 37°C in presence of 5% CO2 .

    Techniques: Purification, Recombinant, Activity Assay, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction