dnase i amplification kit  (Thermo Fisher)


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    Name:
    DNase I Amplification Grade
    Description:
    DNase I Amplification Grade digests single and double stranded DNA to oligodexyribonuleotides containing a 5 phosphate DNase I Amplification Grade is suitable for eliminating DNA during critical RNA purification procedures such as those prior to RNA PCR amplification It is purified and tested for non detectable levels of RNase contamination Absence of RNase is tested by performing a ribonuclease assay with RNA ladder ApplicationRemoving DNA from RNA and protein preparations Specific activitySpecific activity is 10 000 units mg SourcePurified from bovine pancreasPerformance and quality testingRibonuclease assay with RNA ladder and ability to digest single stranded and double stranded DNA to oligonucleotides are determined Unit definitionOne unit increases the absorbance of a high molecular weight DNA solution at a rate of 0 001 A260 units min mL of reaction mixture at 25°C Unit reaction conditions0 1 M sodium acetate pH 5 0 5 mM MgCl2 50 µg mL calf thymus DNA and enzyme in 1 mL for 10 min at 25°C
    Catalog Number:
    18068015
    Price:
    None
    Applications:
    General Real-Time PCR Reagents|PCR & Real-Time PCR|Real Time PCR (qPCR)|Reverse Transcription
    Category:
    Proteins Enzymes Peptides
    Buy from Supplier


    Structured Review

    Thermo Fisher dnase i amplification kit
    DNase I Amplification Grade digests single and double stranded DNA to oligodexyribonuleotides containing a 5 phosphate DNase I Amplification Grade is suitable for eliminating DNA during critical RNA purification procedures such as those prior to RNA PCR amplification It is purified and tested for non detectable levels of RNase contamination Absence of RNase is tested by performing a ribonuclease assay with RNA ladder ApplicationRemoving DNA from RNA and protein preparations Specific activitySpecific activity is 10 000 units mg SourcePurified from bovine pancreasPerformance and quality testingRibonuclease assay with RNA ladder and ability to digest single stranded and double stranded DNA to oligonucleotides are determined Unit definitionOne unit increases the absorbance of a high molecular weight DNA solution at a rate of 0 001 A260 units min mL of reaction mixture at 25°C Unit reaction conditions0 1 M sodium acetate pH 5 0 5 mM MgCl2 50 µg mL calf thymus DNA and enzyme in 1 mL for 10 min at 25°C
    https://www.bioz.com/result/dnase i amplification kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dnase i amplification kit - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Comparing protocols for preparation of DNA-free total yeast RNA suitable for RT-PCR
    Article Snippet: The reaction was stopped by the addition of 2 mM EDTA, pH 8.0, followed by incubation at 37°C for 1 min before inactivation at 65°C for 10 min. RNA was extracted with 1 volume of phenol: chloroform (5:1) and after centrifugation at 2200 × g for 5 min, the RNA present in the upper aqueous phase was precipitated with ice-cold absolute ethanol and collected by centrifugation at 15000 × g for 15 min (4°C). .. Protocol II was performed using the DNase I amplification grade kit (Life Technologies, Inc.) following the recommended procedure.

    Amplification:

    Article Title: Inhibition of the RNA polymerase III-mediated dsDNA-sensing pathway of innate immunity by vaccinia virus protein E3
    Article Snippet: .. Nucleic acids were treated either with RNase A using the conditions described previously ( ) or with amplification grade DNase I following the manufacturer's instructions (Invitrogen). .. Real-time qPCR.

    Article Title: Comparing protocols for preparation of DNA-free total yeast RNA suitable for RT-PCR
    Article Snippet: .. Protocol II was performed using the DNase I amplification grade kit (Life Technologies, Inc.) following the recommended procedure. ..

    Article Title: Allele-specific long-distance regulation dictates IL-32 isoform switching and mediates susceptibility to HIV-1
    Article Snippet: .. Subsequent DNase I digestion was performed with amplification grade DNase I (Invitrogen). .. The reverse-transcriptase reaction was performed using SuperScript II RNase H reverse transcriptase (Invitrogen) according to the manufacturer’s instructions.

    Article Title: Gene expression in isolated plant mitochondria: high fidelity of transcription, splicing and editing of a transgene product in electroporated organelles
    Article Snippet: .. One microgram of RNA was treated with 2 U of amplification grade DNase I (Gibco BRL). cDNA synthesis was performed with 200 U of Superscript II RT using 100 ng of random hexamers as proposed by the supplier. .. The PCR reactions were performed with Advantage™ 2 Polymerase Mix (Clontech) as follows: 95°C for 1 min; 5 cycles at 95°C for 30 s and 68°C for 1 min; 30 cycles at 95°C for 30 s, 58°C for 30 s and 68°C for 30 s; and finally 68°C for 1 min.

    Article Title: Induction of Bex genes by curcumin is associated with apoptosis and activation of p53 in N2a neuroblastoma cells
    Article Snippet: .. Total RNA was treated with 2 units of amplification grade DNase I (Life technologies, USA) for 15 minutes at room temperature. .. The DNase activity was inhibited using EDTA to a final concentration of 2 mM and incubated at 65 °C for 10 minutes.

    Article Title: Immune complexes containing scleroderma-specific autoantibodies induce a profibrotic and proinflammatory phenotype in skin fibroblasts
    Article Snippet: .. Amplification Grade DNase I (ThermoFisher Scientific) was used to eliminate residual genomic DNA. .. A reverse transcription reaction was performed using the SuperScript™ First-Strand Synthesis System for RT-PCR (ThermoFisher Scientific).

    Article Title: C3H Mouse Mammary Tumor Virus Superantigen Function Requires a Splice Donor Site in the Envelope Gene
    Article Snippet: .. Twenty to forty micrograms of total cellular or tissue RNA was DNase treated using 3 to 5 U of amplification grade DNase I (GIBCO BRL) at 37°C for 1 h. DNase I was heat inactivated after the addition of EDTA to a final concentration of 2.5 mM and incubation at 70°C for 10 min. Five to ten micrograms of DNase-treated RNA was used in the reverse transcription (RT) reaction with poly(dT17 ) primer. ..

    Article Title: Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs
    Article Snippet: .. A mixture containing 2 μL of conventional RT mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18 (Thermo Fisher), 8 pmol random hexamers (TaKaRa), and 1.5× PrimeScript enzyme mix in RNase-free water) or 2 μL of RT-RamDA mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18, 8 pmol random hexamers or NSRs, 0.2 U of DNase I Amplification Grade (Thermo Fisher), 100 ng of T4 gene 32 protein (Roche), and 1.5× PrimeScript enzyme mix in RNase-free water) was added to 1 μL of diluted, denatured template RNA. .. The mixture was agitated for 30 s at 2000 rpm using MixMate (Eppendorf) and incubated in a C1000 thermal cycler (Bio-Rad) at 25 °C for 10 min, 30 °C for 10 min, 37 °C for 30 min, 50 °C for 5 min, and 85 °C for 5 min.

    Article Title: Mutation of the Streptococcus gordonii Thiol-Disulfide Oxidoreductase SdbA Leads to Enhanced Biofilm Formation Mediated by the CiaRH Two-Component Signaling System
    Article Snippet: .. The RNA (1 μg) was treated with 1 unit of amplification grade DNase I (Life Technologies Inc., Waltham, MA) for 15 min at room temperature, and removal of DNA was confirmed by PCR using the 16S rRNA primers (SL525/697). cDNA synthesis was carried out using random primers and SuperScript II reverse transcriptase (Life Technologies Inc.) according to the manufacturer’s directions. .. Biofilms for RNA isolation were grown in BM as described above.

    Article Title: Catalytic-Independent Functions of PARP-1 Determine Sox2 Pioneer Activity at Intractable Genomic Loci
    Article Snippet: .. The DNA was digested by the addition of 0.1 unit (for naked DNA) or 1 unit (for nucleosomal DNA) of amplification grade DNase I (Invitrogen) for 5 min. at 25°C. .. Digestion was stopped by addition of an equal volume of DNaseI stop solution containing 20 mM Tris-HCl pH 7.5, 50 mM EDTA, 2% SDS, 0.2 mg/ml proteinase K, 300 ng/μL glycogen with incubation for 1 hour at 55°C.

    Expressing:

    Article Title: Allele-specific long-distance regulation dictates IL-32 isoform switching and mediates susceptibility to HIV-1
    Article Snippet: Subsequent DNase I digestion was performed with amplification grade DNase I (Invitrogen). .. Quantitative real-time PCR reactions for gene expression analysis were performed on a CFX96TM Real-Time System (Bio-Rad) with the GoTaq qPCR Master Mix (Promega) using the following parameters: denaturation at 95°C for 5 min, followed by 40 cycles at 95°C for 10 s, and 60°C for 30 s, followed by a melting curve analysis.

    Article Title: Immune complexes containing scleroderma-specific autoantibodies induce a profibrotic and proinflammatory phenotype in skin fibroblasts
    Article Snippet: Paragraph title: tlr2 , tlr3 , tlr4 , tlr7 , tlr8 , tlr9 , interferon-α , interferon-β , endothelin-1 , collagenIα1 and mmp-1 mRNA expression levels ... Amplification Grade DNase I (ThermoFisher Scientific) was used to eliminate residual genomic DNA.

    Article Title: Mutation of the Streptococcus gordonii Thiol-Disulfide Oxidoreductase SdbA Leads to Enhanced Biofilm Formation Mediated by the CiaRH Two-Component Signaling System
    Article Snippet: Quantitative real-time PCR (qPCR) Expression of ciaR and degP was measured in the biofilm inoculum and in biofilm cells. .. The RNA (1 μg) was treated with 1 unit of amplification grade DNase I (Life Technologies Inc., Waltham, MA) for 15 min at room temperature, and removal of DNA was confirmed by PCR using the 16S rRNA primers (SL525/697). cDNA synthesis was carried out using random primers and SuperScript II reverse transcriptase (Life Technologies Inc.) according to the manufacturer’s directions.

    Modification:

    Article Title: Comparing protocols for preparation of DNA-free total yeast RNA suitable for RT-PCR
    Article Snippet: Protocol I was a modification of the procedure previously described for C. Botulinum [ ]. .. Protocol II was performed using the DNase I amplification grade kit (Life Technologies, Inc.) following the recommended procedure.

    Article Title: Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs
    Article Snippet: The reaction buffer for RT was modified using the PrimeScript RT reagent Kit (TaKaRa). .. A mixture containing 2 μL of conventional RT mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18 (Thermo Fisher), 8 pmol random hexamers (TaKaRa), and 1.5× PrimeScript enzyme mix in RNase-free water) or 2 μL of RT-RamDA mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18, 8 pmol random hexamers or NSRs, 0.2 U of DNase I Amplification Grade (Thermo Fisher), 100 ng of T4 gene 32 protein (Roche), and 1.5× PrimeScript enzyme mix in RNase-free water) was added to 1 μL of diluted, denatured template RNA.

    Concentration Assay:

    Article Title: Induction of Bex genes by curcumin is associated with apoptosis and activation of p53 in N2a neuroblastoma cells
    Article Snippet: Total RNA was treated with 2 units of amplification grade DNase I (Life technologies, USA) for 15 minutes at room temperature. .. The DNase activity was inhibited using EDTA to a final concentration of 2 mM and incubated at 65 °C for 10 minutes.

    Article Title: C3H Mouse Mammary Tumor Virus Superantigen Function Requires a Splice Donor Site in the Envelope Gene
    Article Snippet: .. Twenty to forty micrograms of total cellular or tissue RNA was DNase treated using 3 to 5 U of amplification grade DNase I (GIBCO BRL) at 37°C for 1 h. DNase I was heat inactivated after the addition of EDTA to a final concentration of 2.5 mM and incubation at 70°C for 10 min. Five to ten micrograms of DNase-treated RNA was used in the reverse transcription (RT) reaction with poly(dT17 ) primer. ..

    Mutagenesis:

    Article Title: Mutation of the Streptococcus gordonii Thiol-Disulfide Oxidoreductase SdbA Leads to Enhanced Biofilm Formation Mediated by the CiaRH Two-Component Signaling System
    Article Snippet: For the inoculum, RNA was isolated from cultures of the parent, ΔsdbA , and sdbA- complemented mutant grown in HTVG for 12 h to an OD600 of ~1.0–1.2. .. The RNA (1 μg) was treated with 1 unit of amplification grade DNase I (Life Technologies Inc., Waltham, MA) for 15 min at room temperature, and removal of DNA was confirmed by PCR using the 16S rRNA primers (SL525/697). cDNA synthesis was carried out using random primers and SuperScript II reverse transcriptase (Life Technologies Inc.) according to the manufacturer’s directions.

    Isolation:

    Article Title: Inhibition of the RNA polymerase III-mediated dsDNA-sensing pathway of innate immunity by vaccinia virus protein E3
    Article Snippet: Paragraph title: RNA isolation and enzyme treatment of nucleic acids. ... Nucleic acids were treated either with RNase A using the conditions described previously ( ) or with amplification grade DNase I following the manufacturer's instructions (Invitrogen).

    Article Title: Allele-specific long-distance regulation dictates IL-32 isoform switching and mediates susceptibility to HIV-1
    Article Snippet: cDNA analysis Total cellular RNA was isolated from the different cell lines with TriPure Isolation Reagent according to the manufacturer’s instructions (Roche Diagnostics). .. Subsequent DNase I digestion was performed with amplification grade DNase I (Invitrogen).

    Article Title: Induction of Bex genes by curcumin is associated with apoptosis and activation of p53 in N2a neuroblastoma cells
    Article Snippet: Paragraph title: RNA Isolation ... Total RNA was treated with 2 units of amplification grade DNase I (Life technologies, USA) for 15 minutes at room temperature.

    Article Title: Mutation of the Streptococcus gordonii Thiol-Disulfide Oxidoreductase SdbA Leads to Enhanced Biofilm Formation Mediated by the CiaRH Two-Component Signaling System
    Article Snippet: Total RNA was isolated using the hot acid phenol method, as described previously [ ]. .. The RNA (1 μg) was treated with 1 unit of amplification grade DNase I (Life Technologies Inc., Waltham, MA) for 15 min at room temperature, and removal of DNA was confirmed by PCR using the 16S rRNA primers (SL525/697). cDNA synthesis was carried out using random primers and SuperScript II reverse transcriptase (Life Technologies Inc.) according to the manufacturer’s directions.

    Footprinting:

    Article Title: Catalytic-Independent Functions of PARP-1 Determine Sox2 Pioneer Activity at Intractable Genomic Loci
    Article Snippet: Paragraph title: Footprinting with trinucleosomes ... The DNA was digested by the addition of 0.1 unit (for naked DNA) or 1 unit (for nucleosomal DNA) of amplification grade DNase I (Invitrogen) for 5 min. at 25°C.

    Purification:

    Article Title: Immune complexes containing scleroderma-specific autoantibodies induce a profibrotic and proinflammatory phenotype in skin fibroblasts
    Article Snippet: tlr2 , tlr3 , tlr4 , tlr7 , tlr8 , tlr9 , interferon-α , interferon-β , endothelin-1 , collagenIα1 and mmp-1 mRNA expression levels Total RNA from fibroblasts was purified using Trizol Reagent (ThermoFisher Scientific). .. Amplification Grade DNase I (ThermoFisher Scientific) was used to eliminate residual genomic DNA.

    Article Title: Catalytic-Independent Functions of PARP-1 Determine Sox2 Pioneer Activity at Intractable Genomic Loci
    Article Snippet: For these assays (see ), 46 ng of the three NPE cassette as naked DNA or reconstituted into trinucleosomes was incubated with 25 nM of Sox2 protein (purchased from Abcam; ab169843) in the presence or absence of 10 nM of purified PARP-1 protein in a 20 μL reaction for 1 hour at 30°C under the following buffer conditions: 15 mM Tris-HCl pH 7.5, 0.3 mM EDTA, 0.2 mM DTT, 2% glycerol, 50 mM NaCl, 150 ng/μL BSA, 1x protease inhibitor cocktail (Roche). .. The DNA was digested by the addition of 0.1 unit (for naked DNA) or 1 unit (for nucleosomal DNA) of amplification grade DNase I (Invitrogen) for 5 min. at 25°C.

    Spectrophotometry:

    Article Title: Induction of Bex genes by curcumin is associated with apoptosis and activation of p53 in N2a neuroblastoma cells
    Article Snippet: Total RNA was isolated using Trizol (Invitrogen) as per manufacturer’s instructions and quantified by utilizing Nanovue spectrophotometer (GE healthcare). .. Total RNA was treated with 2 units of amplification grade DNase I (Life technologies, USA) for 15 minutes at room temperature.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Gene expression in isolated plant mitochondria: high fidelity of transcription, splicing and editing of a transgene product in electroporated organelles
    Article Snippet: Paragraph title: RT–PCR ... One microgram of RNA was treated with 2 U of amplification grade DNase I (Gibco BRL). cDNA synthesis was performed with 200 U of Superscript II RT using 100 ng of random hexamers as proposed by the supplier.

    Article Title: Immune complexes containing scleroderma-specific autoantibodies induce a profibrotic and proinflammatory phenotype in skin fibroblasts
    Article Snippet: Amplification Grade DNase I (ThermoFisher Scientific) was used to eliminate residual genomic DNA. .. A reverse transcription reaction was performed using the SuperScript™ First-Strand Synthesis System for RT-PCR (ThermoFisher Scientific).

    Article Title: C3H Mouse Mammary Tumor Virus Superantigen Function Requires a Splice Donor Site in the Envelope Gene
    Article Snippet: Paragraph title: RT-PCR. ... Twenty to forty micrograms of total cellular or tissue RNA was DNase treated using 3 to 5 U of amplification grade DNase I (GIBCO BRL) at 37°C for 1 h. DNase I was heat inactivated after the addition of EDTA to a final concentration of 2.5 mM and incubation at 70°C for 10 min. Five to ten micrograms of DNase-treated RNA was used in the reverse transcription (RT) reaction with poly(dT17 ) primer.

    Real-time Polymerase Chain Reaction:

    Article Title: Allele-specific long-distance regulation dictates IL-32 isoform switching and mediates susceptibility to HIV-1
    Article Snippet: Subsequent DNase I digestion was performed with amplification grade DNase I (Invitrogen). .. Quantitative real-time PCR reactions for gene expression analysis were performed on a CFX96TM Real-Time System (Bio-Rad) with the GoTaq qPCR Master Mix (Promega) using the following parameters: denaturation at 95°C for 5 min, followed by 40 cycles at 95°C for 10 s, and 60°C for 30 s, followed by a melting curve analysis.

    Article Title: Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs
    Article Snippet: A mixture containing 2 μL of conventional RT mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18 (Thermo Fisher), 8 pmol random hexamers (TaKaRa), and 1.5× PrimeScript enzyme mix in RNase-free water) or 2 μL of RT-RamDA mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18, 8 pmol random hexamers or NSRs, 0.2 U of DNase I Amplification Grade (Thermo Fisher), 100 ng of T4 gene 32 protein (Roche), and 1.5× PrimeScript enzyme mix in RNase-free water) was added to 1 μL of diluted, denatured template RNA. .. The RT product was diluted 1:9 in nuclease-free water and used for qPCR analysis.

    Article Title: Mutation of the Streptococcus gordonii Thiol-Disulfide Oxidoreductase SdbA Leads to Enhanced Biofilm Formation Mediated by the CiaRH Two-Component Signaling System
    Article Snippet: Paragraph title: Quantitative real-time PCR (qPCR) ... The RNA (1 μg) was treated with 1 unit of amplification grade DNase I (Life Technologies Inc., Waltham, MA) for 15 min at room temperature, and removal of DNA was confirmed by PCR using the 16S rRNA primers (SL525/697). cDNA synthesis was carried out using random primers and SuperScript II reverse transcriptase (Life Technologies Inc.) according to the manufacturer’s directions.

    Sequencing:

    Article Title: Immune complexes containing scleroderma-specific autoantibodies induce a profibrotic and proinflammatory phenotype in skin fibroblasts
    Article Snippet: Amplification Grade DNase I (ThermoFisher Scientific) was used to eliminate residual genomic DNA. .. Universal PCR Master Mix No AmpErase UNG (ThermoFisher Scientific) was used for quantitative RT-PCR by the ABIPRISM 7900 HT Sequence Detection System (ThermoFisher Scientific).

    Incubation:

    Article Title: Comparing protocols for preparation of DNA-free total yeast RNA suitable for RT-PCR
    Article Snippet: The reaction was stopped by the addition of 2 mM EDTA, pH 8.0, followed by incubation at 37°C for 1 min before inactivation at 65°C for 10 min. RNA was extracted with 1 volume of phenol: chloroform (5:1) and after centrifugation at 2200 × g for 5 min, the RNA present in the upper aqueous phase was precipitated with ice-cold absolute ethanol and collected by centrifugation at 15000 × g for 15 min (4°C). .. Protocol II was performed using the DNase I amplification grade kit (Life Technologies, Inc.) following the recommended procedure.

    Article Title: Induction of Bex genes by curcumin is associated with apoptosis and activation of p53 in N2a neuroblastoma cells
    Article Snippet: Total RNA was treated with 2 units of amplification grade DNase I (Life technologies, USA) for 15 minutes at room temperature. .. The DNase activity was inhibited using EDTA to a final concentration of 2 mM and incubated at 65 °C for 10 minutes.

    Article Title: C3H Mouse Mammary Tumor Virus Superantigen Function Requires a Splice Donor Site in the Envelope Gene
    Article Snippet: .. Twenty to forty micrograms of total cellular or tissue RNA was DNase treated using 3 to 5 U of amplification grade DNase I (GIBCO BRL) at 37°C for 1 h. DNase I was heat inactivated after the addition of EDTA to a final concentration of 2.5 mM and incubation at 70°C for 10 min. Five to ten micrograms of DNase-treated RNA was used in the reverse transcription (RT) reaction with poly(dT17 ) primer. ..

    Article Title: Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs
    Article Snippet: A mixture containing 2 μL of conventional RT mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18 (Thermo Fisher), 8 pmol random hexamers (TaKaRa), and 1.5× PrimeScript enzyme mix in RNase-free water) or 2 μL of RT-RamDA mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18, 8 pmol random hexamers or NSRs, 0.2 U of DNase I Amplification Grade (Thermo Fisher), 100 ng of T4 gene 32 protein (Roche), and 1.5× PrimeScript enzyme mix in RNase-free water) was added to 1 μL of diluted, denatured template RNA. .. The mixture was agitated for 30 s at 2000 rpm using MixMate (Eppendorf) and incubated in a C1000 thermal cycler (Bio-Rad) at 25 °C for 10 min, 30 °C for 10 min, 37 °C for 30 min, 50 °C for 5 min, and 85 °C for 5 min.

    Article Title: Mutation of the Streptococcus gordonii Thiol-Disulfide Oxidoreductase SdbA Leads to Enhanced Biofilm Formation Mediated by the CiaRH Two-Component Signaling System
    Article Snippet: The RNA (1 μg) was treated with 1 unit of amplification grade DNase I (Life Technologies Inc., Waltham, MA) for 15 min at room temperature, and removal of DNA was confirmed by PCR using the 16S rRNA primers (SL525/697). cDNA synthesis was carried out using random primers and SuperScript II reverse transcriptase (Life Technologies Inc.) according to the manufacturer’s directions. .. The plates were incubated at 37°C, 5% CO2 for 24 h. Planktonic cells and spent medium were then removed by pipetting and the biofilms were washed once with 10 ml phosphate buffered saline (PBS).

    Article Title: Catalytic-Independent Functions of PARP-1 Determine Sox2 Pioneer Activity at Intractable Genomic Loci
    Article Snippet: For these assays (see ), 46 ng of the three NPE cassette as naked DNA or reconstituted into trinucleosomes was incubated with 25 nM of Sox2 protein (purchased from Abcam; ab169843) in the presence or absence of 10 nM of purified PARP-1 protein in a 20 μL reaction for 1 hour at 30°C under the following buffer conditions: 15 mM Tris-HCl pH 7.5, 0.3 mM EDTA, 0.2 mM DTT, 2% glycerol, 50 mM NaCl, 150 ng/μL BSA, 1x protease inhibitor cocktail (Roche). .. The DNA was digested by the addition of 0.1 unit (for naked DNA) or 1 unit (for nucleosomal DNA) of amplification grade DNase I (Invitrogen) for 5 min. at 25°C.

    Activity Assay:

    Article Title: Induction of Bex genes by curcumin is associated with apoptosis and activation of p53 in N2a neuroblastoma cells
    Article Snippet: Total RNA was treated with 2 units of amplification grade DNase I (Life technologies, USA) for 15 minutes at room temperature. .. The DNase activity was inhibited using EDTA to a final concentration of 2 mM and incubated at 65 °C for 10 minutes.

    Quantitative RT-PCR:

    Article Title: Immune complexes containing scleroderma-specific autoantibodies induce a profibrotic and proinflammatory phenotype in skin fibroblasts
    Article Snippet: Amplification Grade DNase I (ThermoFisher Scientific) was used to eliminate residual genomic DNA. .. Universal PCR Master Mix No AmpErase UNG (ThermoFisher Scientific) was used for quantitative RT-PCR by the ABIPRISM 7900 HT Sequence Detection System (ThermoFisher Scientific).

    Polyacrylamide Gel Electrophoresis:

    Article Title: Catalytic-Independent Functions of PARP-1 Determine Sox2 Pioneer Activity at Intractable Genomic Loci
    Article Snippet: The DNA was digested by the addition of 0.1 unit (for naked DNA) or 1 unit (for nucleosomal DNA) of amplification grade DNase I (Invitrogen) for 5 min. at 25°C. .. The resulting DNA fragments were run on an 8% PAGE-urea buffer-gradient gel in TBE.

    Protease Inhibitor:

    Article Title: Catalytic-Independent Functions of PARP-1 Determine Sox2 Pioneer Activity at Intractable Genomic Loci
    Article Snippet: For these assays (see ), 46 ng of the three NPE cassette as naked DNA or reconstituted into trinucleosomes was incubated with 25 nM of Sox2 protein (purchased from Abcam; ab169843) in the presence or absence of 10 nM of purified PARP-1 protein in a 20 μL reaction for 1 hour at 30°C under the following buffer conditions: 15 mM Tris-HCl pH 7.5, 0.3 mM EDTA, 0.2 mM DTT, 2% glycerol, 50 mM NaCl, 150 ng/μL BSA, 1x protease inhibitor cocktail (Roche). .. The DNA was digested by the addition of 0.1 unit (for naked DNA) or 1 unit (for nucleosomal DNA) of amplification grade DNase I (Invitrogen) for 5 min. at 25°C.

    Polymerase Chain Reaction:

    Article Title: Gene expression in isolated plant mitochondria: high fidelity of transcription, splicing and editing of a transgene product in electroporated organelles
    Article Snippet: One microgram of RNA was treated with 2 U of amplification grade DNase I (Gibco BRL). cDNA synthesis was performed with 200 U of Superscript II RT using 100 ng of random hexamers as proposed by the supplier. .. The PCR reactions were performed with Advantage™ 2 Polymerase Mix (Clontech) as follows: 95°C for 1 min; 5 cycles at 95°C for 30 s and 68°C for 1 min; 30 cycles at 95°C for 30 s, 58°C for 30 s and 68°C for 30 s; and finally 68°C for 1 min.

    Article Title: Immune complexes containing scleroderma-specific autoantibodies induce a profibrotic and proinflammatory phenotype in skin fibroblasts
    Article Snippet: Amplification Grade DNase I (ThermoFisher Scientific) was used to eliminate residual genomic DNA. .. Universal PCR Master Mix No AmpErase UNG (ThermoFisher Scientific) was used for quantitative RT-PCR by the ABIPRISM 7900 HT Sequence Detection System (ThermoFisher Scientific).

    Article Title: Mutation of the Streptococcus gordonii Thiol-Disulfide Oxidoreductase SdbA Leads to Enhanced Biofilm Formation Mediated by the CiaRH Two-Component Signaling System
    Article Snippet: .. The RNA (1 μg) was treated with 1 unit of amplification grade DNase I (Life Technologies Inc., Waltham, MA) for 15 min at room temperature, and removal of DNA was confirmed by PCR using the 16S rRNA primers (SL525/697). cDNA synthesis was carried out using random primers and SuperScript II reverse transcriptase (Life Technologies Inc.) according to the manufacturer’s directions. .. Biofilms for RNA isolation were grown in BM as described above.

    Lysis:

    Article Title: Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs
    Article Snippet: RT-RamDA Template RNA was diluted in 1 μL of lysis buffer, denatured for 1.5 min at 70 °C, and stored on ice. .. A mixture containing 2 μL of conventional RT mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18 (Thermo Fisher), 8 pmol random hexamers (TaKaRa), and 1.5× PrimeScript enzyme mix in RNase-free water) or 2 μL of RT-RamDA mix (1.5× PrimeScript buffer, 0.6 pmol oligo(dT)18, 8 pmol random hexamers or NSRs, 0.2 U of DNase I Amplification Grade (Thermo Fisher), 100 ng of T4 gene 32 protein (Roche), and 1.5× PrimeScript enzyme mix in RNase-free water) was added to 1 μL of diluted, denatured template RNA.

    Binding Assay:

    Article Title: Catalytic-Independent Functions of PARP-1 Determine Sox2 Pioneer Activity at Intractable Genomic Loci
    Article Snippet: Sox2 binding to naked DNA or the reconstituted trinucleosome templates described above was assayed by DNase I footprinting. .. The DNA was digested by the addition of 0.1 unit (for naked DNA) or 1 unit (for nucleosomal DNA) of amplification grade DNase I (Invitrogen) for 5 min. at 25°C.

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  • 99
    Thermo Fisher dnase amplification grade i kit
    Dnase Amplification Grade I Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnase amplification grade i kit/product/Thermo Fisher
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    dnase amplification grade i kit - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

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