dnase i rnase free  (New England Biolabs)


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    Name:
    DNase I RNase free
    Description:
    DNase I RNase free 5 000 units
    Catalog Number:
    m0303l
    Price:
    276
    Size:
    5 000 units
    Category:
    Deoxyribonucleases DNase
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    Structured Review

    New England Biolabs dnase i rnase free
    DNase I RNase free
    DNase I RNase free 5 000 units
    https://www.bioz.com/result/dnase i rnase free/product/New England Biolabs
    Average 90 stars, based on 50 article reviews
    Price from $9.99 to $1999.99
    dnase i rnase free - by Bioz Stars, 2020-01
    90/100 stars

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    Related Articles

    Centrifugation:

    Article Title: The DNAJA2 Substrate Release Mechanism Is Essential for Chaperone-mediated Folding *
    Article Snippet: Cell samples, taken before heat shock and at 0, 1, and 2 h of recovery, were lysed on ice with PBS containing 1% Triton X-100, and the insoluble material was removed by centrifugation at 20,000 × g . .. The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS.

    Article Title: Extrachloroplastic PP7L Functions in Chloroplast Development and Abiotic Stress Tolerance 1Extrachloroplastic PP7L Functions in Chloroplast Development and Abiotic Stress Tolerance 1 [OPEN]
    Article Snippet: After centrifugation, DNA was precipitated from the supernatant by adding isopropyl alcohol. .. RNA samples were treated with DNase I (New England BioLabs) and quantified with a Nanodrop spectrophotometer (Thermo Fisher Scientific).

    Luciferase:

    Article Title: The DNAJA2 Substrate Release Mechanism Is Essential for Chaperone-mediated Folding *
    Article Snippet: Paragraph title: Luciferase Refolding in Cells ... The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS.

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. PLuc plasmid prib(+)RLuc* 10 U/μL Mlu I (New England BioLabs) 10× NEBuffer 3 (provided with Mlu I) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Phenol:chloroform:IAA (25:24:1) Chloroform:IAA (24:1) 3 M NaOAc (pH 5.2) Isopropanol 75% (v/v) EtOH T7 RNA Polymerase** 5× Transcription Buffer** 40 U/μL RNaseOUT (Invitrogen) NTP Mix (25 mM each NTP) 2U/μL DNase I (RNase-free; New England BioLabs) 3× LiCl-EDTA 0.5× TE 2× RNA Denaturing Loading Dye HeLa or HeLa S3 Cells (Unit 15H.1, Support Protocol 2) D-PBS (PBS w/o Ca2+ or Mg2+ ) 0.05% Trypsin-EDTA 10% NCS DMEM/F-12 2M GuHCl 6-well plates 5× Cell Culture Lysis Reagent (Promega) Luciferase Assay System (Promega) 37ºC heat block or water bath 95ºC heat block Ice NanoDrop spectrophotometer 37ºC and 5% CO2 humidified incubator Electroporator and 4 mm cuvettes Tissue culture microscope (inverted phase-contrast) ..

    Reporter Assay:

    Article Title: The DNAJA2 Substrate Release Mechanism Is Essential for Chaperone-mediated Folding *
    Article Snippet: The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS. .. Luciferase enzymatic activities in the supernatants were measured using the luciferase reporter assay kit (Promega), and activity values were normalized to total protein amounts.

    High Throughput Screening Assay:

    Article Title: A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing
    Article Snippet: Paragraph title: High-throughput Odorant Screening ... Then 0.5 uL DNase I (New England Biolabs) was added to lysate, and incubated at 37°C for 15 minutes.

    Synthesized:

    Article Title: Tomato PEPR1 ORTHOLOG RECEPTOR-LIKE KINASE1 Regulates Responses to Systemin, Necrotrophic Fungi, and Insect Herbivory
    Article Snippet: After extraction, RNA is treated with DNase I (New England Biolabs). .. The first-strand cDNA was synthesized from 2 µg of total RNA using AMV reverse transcriptase (New England Biolabs).

    Blocking Assay:

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. PLuc plasmid prib(+)RLuc* 10 U/μL Mlu I (New England BioLabs) 10× NEBuffer 3 (provided with Mlu I) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Phenol:chloroform:IAA (25:24:1) Chloroform:IAA (24:1) 3 M NaOAc (pH 5.2) Isopropanol 75% (v/v) EtOH T7 RNA Polymerase** 5× Transcription Buffer** 40 U/μL RNaseOUT (Invitrogen) NTP Mix (25 mM each NTP) 2U/μL DNase I (RNase-free; New England BioLabs) 3× LiCl-EDTA 0.5× TE 2× RNA Denaturing Loading Dye HeLa or HeLa S3 Cells (Unit 15H.1, Support Protocol 2) D-PBS (PBS w/o Ca2+ or Mg2+ ) 0.05% Trypsin-EDTA 10% NCS DMEM/F-12 2M GuHCl 6-well plates 5× Cell Culture Lysis Reagent (Promega) Luciferase Assay System (Promega) 37ºC heat block or water bath 95ºC heat block Ice NanoDrop spectrophotometer 37ºC and 5% CO2 humidified incubator Electroporator and 4 mm cuvettes Tissue culture microscope (inverted phase-contrast) ..

    Real-time Polymerase Chain Reaction:

    Article Title: HLA-E expression and its clinical relevance in human renal cell carcinoma
    Article Snippet: .. RNA extraction, cDNA synthesis and qPCR Total RNA was extracted from cell pellets employing TRIzol Reagent (Life Technologies), digested with DNase I (NEB, Ipswich, MA, USA) and applied for cDNA synthesis utilizing the RevertAidTM H Minus First Strand cDNA synthesis kit (Life Technologies) as recently reported in Jasinski-Bergner and co-authors [ ]. ..

    Article Title: A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing
    Article Snippet: Then 0.5 uL DNase I (New England Biolabs) was added to lysate, and incubated at 37°C for 15 minutes. .. Reactions were incubated at 42°C for 60 min and the RT enzyme was heat inactivated at 85°C for 10 min. For each batch, qPCR was performed on a few wells (OL005F and OL013) with SYBR FAST qPCR Mastermix to determine the number of cycles necessary for PCR based library preparation.

    Article Title: Tomato PEPR1 ORTHOLOG RECEPTOR-LIKE KINASE1 Regulates Responses to Systemin, Necrotrophic Fungi, and Insect Herbivory
    Article Snippet: After extraction, RNA is treated with DNase I (New England Biolabs). .. RT-qPCR analysis was performed on Mx3000P real-time PCR detection system (Stratagene) using iTaq Universal SYBR Green Supermix (Bio-Rad).

    Incubation:

    Article Title: A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing
    Article Snippet: .. Then 0.5 uL DNase I (New England Biolabs) was added to lysate, and incubated at 37°C for 15 minutes. ..

    Article Title: In vitro assembly and activity of an archaeal CRISPR-Cas type I-A Cascade interference complex
    Article Snippet: .. The specification of the contaminated nucleic acid was conducted by adding 2 U DNase I (RNase-free, NEB), 1 ng RNase A (biochemistry grade, Ambion) or 25 U benzonase (NEB) in the appropriate reaction buffer to the protein solution and incubated for 30 min at 37°C before loading on agarose gels. .. Transmission electron microscopy (TEM) To further analyze samples of Cas7 purifications, 5 µl of a concentrated protein solution was applied to carbon-coated 400 mesh copper grids.

    Article Title: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
    Article Snippet: .. For DNase I treatment, isolated NETs were incubated with DNase I (NE Biolabs) at 10 U ml−1 for 1 h before addition to SMCs. .. Live imaging of cell death was performed on SMCs stained with Calcein AM (Thermo Fisher, 1:1,000).

    Article Title: Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence
    Article Snippet: .. Nucleic acid was incubated with 5 μL DNase I buffer, 3 μL DNase I (New England Biolabs) and 1 μL Super RNaseIN (Promega) at 37°C for 1 hr. .. Cleanup of RNA was performed with a Qiagen RNeasy mini kit as per manufacturer’s instructions.

    Article Title: Antibiotics Stimulate Formation of Vesicles in Staphylococcus aureus in both Phage-Dependent and -Independent Fashions and via Different Routes
    Article Snippet: .. Briefly, CMVs were resuspended in phosphate-buffered saline (PBS) and were incubated for 1 h in the presence of DNase I (NEB) to degrade extravesicular DNA. .. The DNase was then inactivated at 75C° for 15 min. DNA was released from CMVs by lysis, using 0.125% Triton X-100.

    Article Title: RNA-mediated dimerization of the human deoxycytidine deaminase APOBEC3H influences enzyme activity and interaction with nucleic acids
    Article Snippet: Then the sample was or was not treated with RNase A (Roche Applied Science) or DNase I (New England Biolabs) for 15 min at room temperature. .. For RNase A treatments in the presence of a 118 nt ssDNA oligonucleotide, 3.5 µg (10 μM) A3H was incubated with 10, 50, or 100 μM DNA for 3 min at room temperature before RNase A treatment.

    Proliferation Assay:

    Article Title: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
    Article Snippet: In some experiments, SMC viability was measured using Vybrant MTT cell proliferation assay (Thermo Fisher) according to the manufacturer’s instructions and measured with a plate reader (Tecan, InfiniteF200Pro). .. For DNase I treatment, isolated NETs were incubated with DNase I (NE Biolabs) at 10 U ml−1 for 1 h before addition to SMCs.

    Expressing:

    Article Title: Tomato PEPR1 ORTHOLOG RECEPTOR-LIKE KINASE1 Regulates Responses to Systemin, Necrotrophic Fungi, and Insect Herbivory
    Article Snippet: Paragraph title: RNA Extraction and Analysis of Gene Expression ... After extraction, RNA is treated with DNase I (New England Biolabs).

    BIA-KA:

    Article Title: The DNAJA2 Substrate Release Mechanism Is Essential for Chaperone-mediated Folding *
    Article Snippet: The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS. .. The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS.

    Western Blot:

    Article Title: The DNAJA2 Substrate Release Mechanism Is Essential for Chaperone-mediated Folding *
    Article Snippet: The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS. .. Equal amounts of protein were analyzed on immunoblots.

    Derivative Assay:

    Article Title: Antibiotics Stimulate Formation of Vesicles in Staphylococcus aureus in both Phage-Dependent and -Independent Fashions and via Different Routes
    Article Snippet: The DNA loads of CMVs derived from strains NRS135 and NRS77phage stimulated with either MMC at 100 ng/ml or antibiotics at 10× MIC were quantified as described previously , with some minor modifications. .. Briefly, CMVs were resuspended in phosphate-buffered saline (PBS) and were incubated for 1 h in the presence of DNase I (NEB) to degrade extravesicular DNA.

    Transfection:

    Article Title: The DNAJA2 Substrate Release Mechanism Is Essential for Chaperone-mediated Folding *
    Article Snippet: One day after transfection, cells were treated with cycloheximide (Sigma) at 50 μg/ml to inhibit protein synthesis, transferred to 45 °C for 1 h, and then brought back to 37 °C for up to 2 h recovery. .. The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS.

    Concentration Assay:

    Article Title: A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing
    Article Snippet: Each plate contained two control odorants at a three concentration (10 uM, 100 uM, 1 mM) in triplicate and three wells containing 1% DMSO dissolved in media. .. Then 0.5 uL DNase I (New England Biolabs) was added to lysate, and incubated at 37°C for 15 minutes.

    Article Title: Distinct 5-methylcytosine profiles in poly(A) RNA from mouse embryonic stem cells and brain
    Article Snippet: RNA was treated with 2U of DNase I (New England Biolabs, Ipswich, MA, USA) for 15 min at 37 °C and purified using RNA Clean & Concentrator 25 Kit (Zymo Research, Irvine, CA, USA). .. RNA quality was assessed using an Agilent 2100 Bioanalyzer, and concentration was determined by measuring absorbance at 260 nm and 280 nm in a UV/vis-spectrophotometer.

    Article Title: Tomato PEPR1 ORTHOLOG RECEPTOR-LIKE KINASE1 Regulates Responses to Systemin, Necrotrophic Fungi, and Insect Herbivory
    Article Snippet: After extraction, RNA is treated with DNase I (New England Biolabs). .. RNA concentration was measured using Nanodrop 2000c (Thermo Scientific).

    Cell Culture:

    Article Title: A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing
    Article Snippet: The library was incubated with odorants for three hours in a cell culture incubator with the lids removed. .. Then 0.5 uL DNase I (New England Biolabs) was added to lysate, and incubated at 37°C for 15 minutes.

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. PLuc plasmid prib(+)RLuc* 10 U/μL Mlu I (New England BioLabs) 10× NEBuffer 3 (provided with Mlu I) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Phenol:chloroform:IAA (25:24:1) Chloroform:IAA (24:1) 3 M NaOAc (pH 5.2) Isopropanol 75% (v/v) EtOH T7 RNA Polymerase** 5× Transcription Buffer** 40 U/μL RNaseOUT (Invitrogen) NTP Mix (25 mM each NTP) 2U/μL DNase I (RNase-free; New England BioLabs) 3× LiCl-EDTA 0.5× TE 2× RNA Denaturing Loading Dye HeLa or HeLa S3 Cells (Unit 15H.1, Support Protocol 2) D-PBS (PBS w/o Ca2+ or Mg2+ ) 0.05% Trypsin-EDTA 10% NCS DMEM/F-12 2M GuHCl 6-well plates 5× Cell Culture Lysis Reagent (Promega) Luciferase Assay System (Promega) 37ºC heat block or water bath 95ºC heat block Ice NanoDrop spectrophotometer 37ºC and 5% CO2 humidified incubator Electroporator and 4 mm cuvettes Tissue culture microscope (inverted phase-contrast) ..

    Generated:

    Article Title: Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence
    Article Snippet: Nucleic acid was incubated with 5 μL DNase I buffer, 3 μL DNase I (New England Biolabs) and 1 μL Super RNaseIN (Promega) at 37°C for 1 hr. .. DNase- and RNase-free reagents were used throughout. cDNA was generated with SmartScribe reverse transcriptase according to the manufacturer’s recommendations (New England Biolabs).

    Inhibition:

    Article Title: The DNAJA2 Substrate Release Mechanism Is Essential for Chaperone-mediated Folding *
    Article Snippet: For proteasome inhibition, MG132 (Sigma) was added during the heat shock and recovery phases at 5 μg/ml. .. The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS.

    Imaging:

    Article Title: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
    Article Snippet: For DNase I treatment, isolated NETs were incubated with DNase I (NE Biolabs) at 10 U ml−1 for 1 h before addition to SMCs. .. Live imaging of cell death was performed on SMCs stained with Calcein AM (Thermo Fisher, 1:1,000).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: HLA-E expression and its clinical relevance in human renal cell carcinoma
    Article Snippet: RNA extraction, cDNA synthesis and qPCR Total RNA was extracted from cell pellets employing TRIzol Reagent (Life Technologies), digested with DNase I (NEB, Ipswich, MA, USA) and applied for cDNA synthesis utilizing the RevertAidTM H Minus First Strand cDNA synthesis kit (Life Technologies) as recently reported in Jasinski-Bergner and co-authors [ ]. .. The qPCR reactions were performed in a Rotorgene cycler (Qiagen, Hilden, Germany) at 95°C for 5 min followed by 40 cycles of 95°C for 30 s, 60°C for 30 s and 72°C for 40 s. HLA-E, HLA-F and HLA-G mRNA detection from biopsies were further characterized by semi-quantitative PCR [ ] using the Titan One Tube RT-PCR System (Roche Diagnostics, Heidelberg, Germany) and specific primers for HLA-G: G.257 (forward: 5′-GGAAGAGGAGACACGGAACA-3′) and G.1225 (reverse: 5′-TGAGACAGAGACGGAGACAT-3′) by Real et al., 1999 [ ], Tm 61°C and resulting PCR products were size fractionated on 1% ethidium bromide containing agarose gels [ ].

    Article Title: Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence
    Article Snippet: Paragraph title: RNA preparation and reverse transcriptase RT-PCR ... Nucleic acid was incubated with 5 μL DNase I buffer, 3 μL DNase I (New England Biolabs) and 1 μL Super RNaseIN (Promega) at 37°C for 1 hr.

    Binding Assay:

    Article Title: In vitro assembly and activity of an archaeal CRISPR-Cas type I-A Cascade interference complex
    Article Snippet: Binding of the two Cas7 versions (Cas7, Cas7-SUMO) to contaminating nucleic acids during the purification process was verified by loading 100 µg protein on 1% TAE-agarose gels stained with ethidium bromide alongside the DNA marker (2-log DNA ladder, NEB). .. The specification of the contaminated nucleic acid was conducted by adding 2 U DNase I (RNase-free, NEB), 1 ng RNase A (biochemistry grade, Ambion) or 25 U benzonase (NEB) in the appropriate reaction buffer to the protein solution and incubated for 30 min at 37°C before loading on agarose gels.

    MTT Assay:

    Article Title: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
    Article Snippet: In some experiments, SMC viability was measured using Vybrant MTT cell proliferation assay (Thermo Fisher) according to the manufacturer’s instructions and measured with a plate reader (Tecan, InfiniteF200Pro). .. For DNase I treatment, isolated NETs were incubated with DNase I (NE Biolabs) at 10 U ml−1 for 1 h before addition to SMCs.

    Fluorescence:

    Article Title: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
    Article Snippet: PI+ cells were visualized using a climate chamber fluorescence microscope (Leica, DMi8) and quantified by ImageJ software. .. For DNase I treatment, isolated NETs were incubated with DNase I (NE Biolabs) at 10 U ml−1 for 1 h before addition to SMCs.

    Isolation:

    Article Title: Distinct 5-methylcytosine profiles in poly(A) RNA from mouse embryonic stem cells and brain
    Article Snippet: Paragraph title: RNA isolation ... RNA was treated with 2U of DNase I (New England Biolabs, Ipswich, MA, USA) for 15 min at 37 °C and purified using RNA Clean & Concentrator 25 Kit (Zymo Research, Irvine, CA, USA).

    Article Title: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
    Article Snippet: .. For DNase I treatment, isolated NETs were incubated with DNase I (NE Biolabs) at 10 U ml−1 for 1 h before addition to SMCs. .. Live imaging of cell death was performed on SMCs stained with Calcein AM (Thermo Fisher, 1:1,000).

    Article Title: Extrachloroplastic PP7L Functions in Chloroplast Development and Abiotic Stress Tolerance 1Extrachloroplastic PP7L Functions in Chloroplast Development and Abiotic Stress Tolerance 1 [OPEN]
    Article Snippet: For RNA isolation, frozen tissue was ground in liquid N. Total RNA was extracted by using TRIzol reagent (Invitrogen) according to the manufacturer’s protocol. .. RNA samples were treated with DNase I (New England BioLabs) and quantified with a Nanodrop spectrophotometer (Thermo Fisher Scientific).

    Article Title: Tomato PEPR1 ORTHOLOG RECEPTOR-LIKE KINASE1 Regulates Responses to Systemin, Necrotrophic Fungi, and Insect Herbivory
    Article Snippet: Total RNA was isolated with Trizol reagent from tomato or Arabidopsis leaf tissues according to the manufacturer’s instructions (Invitrogen). .. After extraction, RNA is treated with DNase I (New England Biolabs).

    Flow Cytometry:

    Article Title: A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing
    Article Snippet: This was because lids of 96-well culture plates still allow air flow between wells and we thought that cross contamination of odorants might occur more readily if the odorants were trapped in the small volume of the plate. .. Then 0.5 uL DNase I (New England Biolabs) was added to lysate, and incubated at 37°C for 15 minutes.

    Microscopy:

    Article Title: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
    Article Snippet: PI+ cells were visualized using a climate chamber fluorescence microscope (Leica, DMi8) and quantified by ImageJ software. .. For DNase I treatment, isolated NETs were incubated with DNase I (NE Biolabs) at 10 U ml−1 for 1 h before addition to SMCs.

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. PLuc plasmid prib(+)RLuc* 10 U/μL Mlu I (New England BioLabs) 10× NEBuffer 3 (provided with Mlu I) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Phenol:chloroform:IAA (25:24:1) Chloroform:IAA (24:1) 3 M NaOAc (pH 5.2) Isopropanol 75% (v/v) EtOH T7 RNA Polymerase** 5× Transcription Buffer** 40 U/μL RNaseOUT (Invitrogen) NTP Mix (25 mM each NTP) 2U/μL DNase I (RNase-free; New England BioLabs) 3× LiCl-EDTA 0.5× TE 2× RNA Denaturing Loading Dye HeLa or HeLa S3 Cells (Unit 15H.1, Support Protocol 2) D-PBS (PBS w/o Ca2+ or Mg2+ ) 0.05% Trypsin-EDTA 10% NCS DMEM/F-12 2M GuHCl 6-well plates 5× Cell Culture Lysis Reagent (Promega) Luciferase Assay System (Promega) 37ºC heat block or water bath 95ºC heat block Ice NanoDrop spectrophotometer 37ºC and 5% CO2 humidified incubator Electroporator and 4 mm cuvettes Tissue culture microscope (inverted phase-contrast) ..

    Purification:

    Article Title: Distinct 5-methylcytosine profiles in poly(A) RNA from mouse embryonic stem cells and brain
    Article Snippet: .. RNA was treated with 2U of DNase I (New England Biolabs, Ipswich, MA, USA) for 15 min at 37 °C and purified using RNA Clean & Concentrator 25 Kit (Zymo Research, Irvine, CA, USA). .. Isolated RNA was then subjected to two rounds of poly(A) RNA enrichment using fresh Dynabeads (Ambion) for each round.

    Article Title: In vitro assembly and activity of an archaeal CRISPR-Cas type I-A Cascade interference complex
    Article Snippet: Binding of the two Cas7 versions (Cas7, Cas7-SUMO) to contaminating nucleic acids during the purification process was verified by loading 100 µg protein on 1% TAE-agarose gels stained with ethidium bromide alongside the DNA marker (2-log DNA ladder, NEB). .. The specification of the contaminated nucleic acid was conducted by adding 2 U DNase I (RNase-free, NEB), 1 ng RNase A (biochemistry grade, Ambion) or 25 U benzonase (NEB) in the appropriate reaction buffer to the protein solution and incubated for 30 min at 37°C before loading on agarose gels.

    Article Title: RNA-mediated dimerization of the human deoxycytidine deaminase APOBEC3H influences enzyme activity and interaction with nucleic acids
    Article Snippet: To examine the nature of the nucleic acids present in purified A3H, the same amount of A3H (3.5 μg for WT, GST-WT, Y112A/Y113A, GST-Y112A/Y113A, GST-W115A, GST-R165E/R176E) was or was not treated with Proteinase K (New England Biolabs) for 15 min at room temperature followed by heat inactivation for 10 min at 95°C. .. Then the sample was or was not treated with RNase A (Roche Applied Science) or DNase I (New England Biolabs) for 15 min at room temperature.

    Polymerase Chain Reaction:

    Article Title: HLA-E expression and its clinical relevance in human renal cell carcinoma
    Article Snippet: RNA extraction, cDNA synthesis and qPCR Total RNA was extracted from cell pellets employing TRIzol Reagent (Life Technologies), digested with DNase I (NEB, Ipswich, MA, USA) and applied for cDNA synthesis utilizing the RevertAidTM H Minus First Strand cDNA synthesis kit (Life Technologies) as recently reported in Jasinski-Bergner and co-authors [ ]. .. The qPCR reactions were performed in a Rotorgene cycler (Qiagen, Hilden, Germany) at 95°C for 5 min followed by 40 cycles of 95°C for 30 s, 60°C for 30 s and 72°C for 40 s. HLA-E, HLA-F and HLA-G mRNA detection from biopsies were further characterized by semi-quantitative PCR [ ] using the Titan One Tube RT-PCR System (Roche Diagnostics, Heidelberg, Germany) and specific primers for HLA-G: G.257 (forward: 5′-GGAAGAGGAGACACGGAACA-3′) and G.1225 (reverse: 5′-TGAGACAGAGACGGAGACAT-3′) by Real et al., 1999 [ ], Tm 61°C and resulting PCR products were size fractionated on 1% ethidium bromide containing agarose gels [ ].

    Article Title: A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing
    Article Snippet: Then 0.5 uL DNase I (New England Biolabs) was added to lysate, and incubated at 37°C for 15 minutes. .. Reactions were incubated at 42°C for 60 min and the RT enzyme was heat inactivated at 85°C for 10 min. For each batch, qPCR was performed on a few wells (OL005F and OL013) with SYBR FAST qPCR Mastermix to determine the number of cycles necessary for PCR based library preparation.

    Article Title: CTCF Regulates Kaposi's Sarcoma-Associated Herpesvirus Latency Transcription by Nucleosome Displacement and RNA Polymerase Programming
    Article Snippet: Total RNA was extracted from KSHV bacmid-transformed 293T cells using an RNeasy kit (Qiagen) and then further treated with DNase I (New England Biolabs). .. For semiquantitative PCR assays, LTc was assayed with the LTc-F and LT-R primer set.

    Quantitative RT-PCR:

    Article Title: Tomato PEPR1 ORTHOLOG RECEPTOR-LIKE KINASE1 Regulates Responses to Systemin, Necrotrophic Fungi, and Insect Herbivory
    Article Snippet: After extraction, RNA is treated with DNase I (New England Biolabs). .. RT-qPCR analysis was performed on Mx3000P real-time PCR detection system (Stratagene) using iTaq Universal SYBR Green Supermix (Bio-Rad).

    Staining:

    Article Title: In vitro assembly and activity of an archaeal CRISPR-Cas type I-A Cascade interference complex
    Article Snippet: Binding of the two Cas7 versions (Cas7, Cas7-SUMO) to contaminating nucleic acids during the purification process was verified by loading 100 µg protein on 1% TAE-agarose gels stained with ethidium bromide alongside the DNA marker (2-log DNA ladder, NEB). .. The specification of the contaminated nucleic acid was conducted by adding 2 U DNase I (RNase-free, NEB), 1 ng RNase A (biochemistry grade, Ambion) or 25 U benzonase (NEB) in the appropriate reaction buffer to the protein solution and incubated for 30 min at 37°C before loading on agarose gels.

    Article Title: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
    Article Snippet: For DNase I treatment, isolated NETs were incubated with DNase I (NE Biolabs) at 10 U ml−1 for 1 h before addition to SMCs. .. Live imaging of cell death was performed on SMCs stained with Calcein AM (Thermo Fisher, 1:1,000).

    Plasmid Preparation:

    Article Title: The DNAJA2 Substrate Release Mechanism Is Essential for Chaperone-mediated Folding *
    Article Snippet: HEK293 cells were transfected with 2 μg of luciferase plasmid and 8 μg of DJA2 plasmid or control vector per 60-mm dish. .. The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS.

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. PLuc plasmid prib(+)RLuc* 10 U/μL Mlu I (New England BioLabs) 10× NEBuffer 3 (provided with Mlu I) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Phenol:chloroform:IAA (25:24:1) Chloroform:IAA (24:1) 3 M NaOAc (pH 5.2) Isopropanol 75% (v/v) EtOH T7 RNA Polymerase** 5× Transcription Buffer** 40 U/μL RNaseOUT (Invitrogen) NTP Mix (25 mM each NTP) 2U/μL DNase I (RNase-free; New England BioLabs) 3× LiCl-EDTA 0.5× TE 2× RNA Denaturing Loading Dye HeLa or HeLa S3 Cells (Unit 15H.1, Support Protocol 2) D-PBS (PBS w/o Ca2+ or Mg2+ ) 0.05% Trypsin-EDTA 10% NCS DMEM/F-12 2M GuHCl 6-well plates 5× Cell Culture Lysis Reagent (Promega) Luciferase Assay System (Promega) 37ºC heat block or water bath 95ºC heat block Ice NanoDrop spectrophotometer 37ºC and 5% CO2 humidified incubator Electroporator and 4 mm cuvettes Tissue culture microscope (inverted phase-contrast) ..

    Software:

    Article Title: Externalized histone H4 orchestrates chronic inflammation by inducing lytic cell death
    Article Snippet: PI+ cells were visualized using a climate chamber fluorescence microscope (Leica, DMi8) and quantified by ImageJ software. .. For DNase I treatment, isolated NETs were incubated with DNase I (NE Biolabs) at 10 U ml−1 for 1 h before addition to SMCs.

    SYBR Green Assay:

    Article Title: Tomato PEPR1 ORTHOLOG RECEPTOR-LIKE KINASE1 Regulates Responses to Systemin, Necrotrophic Fungi, and Insect Herbivory
    Article Snippet: After extraction, RNA is treated with DNase I (New England Biolabs). .. RT-qPCR analysis was performed on Mx3000P real-time PCR detection system (Stratagene) using iTaq Universal SYBR Green Supermix (Bio-Rad).

    RNA Extraction:

    Article Title: HLA-E expression and its clinical relevance in human renal cell carcinoma
    Article Snippet: .. RNA extraction, cDNA synthesis and qPCR Total RNA was extracted from cell pellets employing TRIzol Reagent (Life Technologies), digested with DNase I (NEB, Ipswich, MA, USA) and applied for cDNA synthesis utilizing the RevertAidTM H Minus First Strand cDNA synthesis kit (Life Technologies) as recently reported in Jasinski-Bergner and co-authors [ ]. ..

    Article Title: Tomato PEPR1 ORTHOLOG RECEPTOR-LIKE KINASE1 Regulates Responses to Systemin, Necrotrophic Fungi, and Insect Herbivory
    Article Snippet: Paragraph title: RNA Extraction and Analysis of Gene Expression ... After extraction, RNA is treated with DNase I (New England Biolabs).

    Spectrophotometry:

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. PLuc plasmid prib(+)RLuc* 10 U/μL Mlu I (New England BioLabs) 10× NEBuffer 3 (provided with Mlu I) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Phenol:chloroform:IAA (25:24:1) Chloroform:IAA (24:1) 3 M NaOAc (pH 5.2) Isopropanol 75% (v/v) EtOH T7 RNA Polymerase** 5× Transcription Buffer** 40 U/μL RNaseOUT (Invitrogen) NTP Mix (25 mM each NTP) 2U/μL DNase I (RNase-free; New England BioLabs) 3× LiCl-EDTA 0.5× TE 2× RNA Denaturing Loading Dye HeLa or HeLa S3 Cells (Unit 15H.1, Support Protocol 2) D-PBS (PBS w/o Ca2+ or Mg2+ ) 0.05% Trypsin-EDTA 10% NCS DMEM/F-12 2M GuHCl 6-well plates 5× Cell Culture Lysis Reagent (Promega) Luciferase Assay System (Promega) 37ºC heat block or water bath 95ºC heat block Ice NanoDrop spectrophotometer 37ºC and 5% CO2 humidified incubator Electroporator and 4 mm cuvettes Tissue culture microscope (inverted phase-contrast) ..

    Article Title: Extrachloroplastic PP7L Functions in Chloroplast Development and Abiotic Stress Tolerance 1Extrachloroplastic PP7L Functions in Chloroplast Development and Abiotic Stress Tolerance 1 [OPEN]
    Article Snippet: .. RNA samples were treated with DNase I (New England BioLabs) and quantified with a Nanodrop spectrophotometer (Thermo Fisher Scientific). .. Isolated RNA was stored at −80°C until further use.

    Mobility Shift:

    Article Title: In vitro assembly and activity of an archaeal CRISPR-Cas type I-A Cascade interference complex
    Article Snippet: Paragraph title: Electrophorectic mobility shift assay ... The specification of the contaminated nucleic acid was conducted by adding 2 U DNase I (RNase-free, NEB), 1 ng RNase A (biochemistry grade, Ambion) or 25 U benzonase (NEB) in the appropriate reaction buffer to the protein solution and incubated for 30 min at 37°C before loading on agarose gels.

    Lysis:

    Article Title: A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing
    Article Snippet: After odor incubation, media was pipetted out of the plates and cells were lysed by adding 25 uL of ice-cold Cells-to-cDNA II Lysis Buffer (Thermo Fisher) and pipetting up and down to homogenize and lyse cells. .. Then 0.5 uL DNase I (New England Biolabs) was added to lysate, and incubated at 37°C for 15 minutes.

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. PLuc plasmid prib(+)RLuc* 10 U/μL Mlu I (New England BioLabs) 10× NEBuffer 3 (provided with Mlu I) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Phenol:chloroform:IAA (25:24:1) Chloroform:IAA (24:1) 3 M NaOAc (pH 5.2) Isopropanol 75% (v/v) EtOH T7 RNA Polymerase** 5× Transcription Buffer** 40 U/μL RNaseOUT (Invitrogen) NTP Mix (25 mM each NTP) 2U/μL DNase I (RNase-free; New England BioLabs) 3× LiCl-EDTA 0.5× TE 2× RNA Denaturing Loading Dye HeLa or HeLa S3 Cells (Unit 15H.1, Support Protocol 2) D-PBS (PBS w/o Ca2+ or Mg2+ ) 0.05% Trypsin-EDTA 10% NCS DMEM/F-12 2M GuHCl 6-well plates 5× Cell Culture Lysis Reagent (Promega) Luciferase Assay System (Promega) 37ºC heat block or water bath 95ºC heat block Ice NanoDrop spectrophotometer 37ºC and 5% CO2 humidified incubator Electroporator and 4 mm cuvettes Tissue culture microscope (inverted phase-contrast) ..

    Article Title: Antibiotics Stimulate Formation of Vesicles in Staphylococcus aureus in both Phage-Dependent and -Independent Fashions and via Different Routes
    Article Snippet: Briefly, CMVs were resuspended in phosphate-buffered saline (PBS) and were incubated for 1 h in the presence of DNase I (NEB) to degrade extravesicular DNA. .. The DNase was then inactivated at 75C° for 15 min. DNA was released from CMVs by lysis, using 0.125% Triton X-100.

    Marker:

    Article Title: In vitro assembly and activity of an archaeal CRISPR-Cas type I-A Cascade interference complex
    Article Snippet: Binding of the two Cas7 versions (Cas7, Cas7-SUMO) to contaminating nucleic acids during the purification process was verified by loading 100 µg protein on 1% TAE-agarose gels stained with ethidium bromide alongside the DNA marker (2-log DNA ladder, NEB). .. The specification of the contaminated nucleic acid was conducted by adding 2 U DNase I (RNase-free, NEB), 1 ng RNase A (biochemistry grade, Ambion) or 25 U benzonase (NEB) in the appropriate reaction buffer to the protein solution and incubated for 30 min at 37°C before loading on agarose gels.

    Activity Assay:

    Article Title: The DNAJA2 Substrate Release Mechanism Is Essential for Chaperone-mediated Folding *
    Article Snippet: The pellets were resuspended in PBS containing 0.1% Triton X-100, treated with 2 units of DNase I (New England Biolabs) at 37 °C for 30 min, and then denatured with 2% SDS. .. Luciferase enzymatic activities in the supernatants were measured using the luciferase reporter assay kit (Promega), and activity values were normalized to total protein amounts.

    Variant Assay:

    Article Title: Virus-Inspired Membrane Encapsulation of DNA Nanostructures To Achieve In Vivo Stability
    Article Snippet: .. Ten units of DNase I were added to 3× replicates of each variant, and an equivalent volume of DNase I buffer was added to 3× replicates as negative controls. .. All samples were incubated for 24 h at 37 °C on a Tetrad 2 Peltier thermocycler.

    other:

    Article Title: A Small RNA Transforms the Multidrug Resistance of Pseudomonas aeruginosa to Drug Susceptibility
    Article Snippet: The total RNA was then treated with DNase I (New England Biolabs), and the rRNA was depleted using MICROBExpress Bacteria mRNA Enrichment Kit (Ambion), followed by Ribo-Zero rRNA Removal Kits (Gram-Negative Bacteria, Epicentre), according to the manufacturer’s instructions.

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    New England Biolabs dnase i
    Rad26 binds to the upstream DNA and bubble fork of Pol II EC and bends the upstream DNA a , Atomic model for the scaffold in the Pol II EC-Rad26 complex displayed inside the segmented cryo-EM density, obtained by subtracting Pol II and Rad26 from the Pol II EC-Rad26 map. Orange asterisk: Rad26 density not modeled. Top-right inset: scaffold density (in yellow) in the context of the full complex. b , Superposition of the scaffolds from the Pol II EC-Rad26 and Pol II EC structures, with the latter shown in darker colors. c , Close-up view of the interaction between Rad26 and the scaffold. d , <t>DNase</t> I footprinting assay of Pol II EC-Rad26. The experiment was repeated independently 3 times with similar results. e , Close-up of the Rad26 HD2-1 “wedge” (yellow arrow) that interacts with the upstream bubble fork. Same view as c except with Rad26 in a surface charge representation. Right inset: closer view of the interaction, looking from the transcription bubble towards the upstream DNA. g , Major interactions between Rad26 and the Wall and Protrusion regions of Pol II. The Pol II motifs that bind to Rad26 are shown as surface representations, with the corresponding residues listed. f , Effect of transcription bubble size in the affinity of Rad26 for Pol II EC. Mismatches were added to the upstream fork, downstream fork, or both. Data shown as mean and standard deviation (n = 3). P-values (two-tailed Student’s t test): not shown = not significant; * =
    Dnase I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 374 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Rad26 binds to the upstream DNA and bubble fork of Pol II EC and bends the upstream DNA a , Atomic model for the scaffold in the Pol II EC-Rad26 complex displayed inside the segmented cryo-EM density, obtained by subtracting Pol II and Rad26 from the Pol II EC-Rad26 map. Orange asterisk: Rad26 density not modeled. Top-right inset: scaffold density (in yellow) in the context of the full complex. b , Superposition of the scaffolds from the Pol II EC-Rad26 and Pol II EC structures, with the latter shown in darker colors. c , Close-up view of the interaction between Rad26 and the scaffold. d , DNase I footprinting assay of Pol II EC-Rad26. The experiment was repeated independently 3 times with similar results. e , Close-up of the Rad26 HD2-1 “wedge” (yellow arrow) that interacts with the upstream bubble fork. Same view as c except with Rad26 in a surface charge representation. Right inset: closer view of the interaction, looking from the transcription bubble towards the upstream DNA. g , Major interactions between Rad26 and the Wall and Protrusion regions of Pol II. The Pol II motifs that bind to Rad26 are shown as surface representations, with the corresponding residues listed. f , Effect of transcription bubble size in the affinity of Rad26 for Pol II EC. Mismatches were added to the upstream fork, downstream fork, or both. Data shown as mean and standard deviation (n = 3). P-values (two-tailed Student’s t test): not shown = not significant; * =

    Journal: Nature

    Article Title: Structural Basis for Eukaryotic Transcription-Coupled DNA Repair Initiation

    doi: 10.1038/nature24658

    Figure Lengend Snippet: Rad26 binds to the upstream DNA and bubble fork of Pol II EC and bends the upstream DNA a , Atomic model for the scaffold in the Pol II EC-Rad26 complex displayed inside the segmented cryo-EM density, obtained by subtracting Pol II and Rad26 from the Pol II EC-Rad26 map. Orange asterisk: Rad26 density not modeled. Top-right inset: scaffold density (in yellow) in the context of the full complex. b , Superposition of the scaffolds from the Pol II EC-Rad26 and Pol II EC structures, with the latter shown in darker colors. c , Close-up view of the interaction between Rad26 and the scaffold. d , DNase I footprinting assay of Pol II EC-Rad26. The experiment was repeated independently 3 times with similar results. e , Close-up of the Rad26 HD2-1 “wedge” (yellow arrow) that interacts with the upstream bubble fork. Same view as c except with Rad26 in a surface charge representation. Right inset: closer view of the interaction, looking from the transcription bubble towards the upstream DNA. g , Major interactions between Rad26 and the Wall and Protrusion regions of Pol II. The Pol II motifs that bind to Rad26 are shown as surface representations, with the corresponding residues listed. f , Effect of transcription bubble size in the affinity of Rad26 for Pol II EC. Mismatches were added to the upstream fork, downstream fork, or both. Data shown as mean and standard deviation (n = 3). P-values (two-tailed Student’s t test): not shown = not significant; * =

    Article Snippet: DNase I footprinting An aliquot of 20 nM Pol II EC (with 5´-P labeled template DNA strand) was incubated with 0-150 nM Rad26 in the binding buffer (see above) at 23 °C for 30 min. Then DNase I (NEB, USA) was added to a final concentration of 0.04 units/ml and the digestion was carried out for 1 minute (50 seconds if Rad26 was absent) at 23 °C.

    Techniques: Footprinting, Standard Deviation, Two Tailed Test