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Thermo Fisher mnase i
Mnase I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/mnase i/product/Thermo Fisher
Average 88 stars, based on 1 article reviews
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mnase i - by Bioz Stars, 2020-05
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Article Title: RNA-dependent recruitment of the origin recognition complex
Article Snippet: For micrococcal nuclease I conditions, 0.2 U/ml MNase I (USB Corp.) was added to beads resuspended in MNase I buffer (10 mM Tris–HCl pH 8.0, 1 mM CaCl2 ).

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    Thermo Fisher mnase
    <t>DKC1-associated</t> RNAs in recombinant DKC1 complexes are resistant to extensive <t>MNase</t> digestion. RNAs co-purified from mock and MNase-treated recombinant DKC1 complexes in Figure 4—figure supplement 1 are 5′ end radiolabeled and separated on a 6% urea-polyacrylamide gel as described in Figure 2—figure supplement 1 . Note that the prominently labeled 130–140 nucleotide (nt)-long RNA clusters are resistant to complete nuclease digestion. It appears that these RNAs are cut on average once by MNase to generate two new smaller clusters (80–90 nt and 30–55 nt) that remain stably associated with the DKC1 complexes. Increasing the amount of MNase and/or nuclease digestion time did not change the patterns or disrupt the integrity of the protein complexes (data not shown). DOI: http://dx.doi.org/10.7554/eLife.03573.010
    Mnase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mnase/product/Thermo Fisher
    Average 92 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    mnase - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    93
    Thermo Fisher purification gst mnase
    MapR and RHΔ CUT RUN Signals Are Enriched at Similar Regions Genome-wide (A) Schematic of RHΔC R using FLAG M2 antibody (left) and MapR using <t>GST-RHΔ-MNase</t> (right) in HEK293. (B) Enriched regions identified by RHΔC R and R-ChIP in HEK293. GRO-seq and H3K4me3 tracks indicate active gene transcription. (C) Venn diagram of gene-level overlap between RHΔC R and R-ChIP. Total number of unique genes with an R-loop at the promoter region (−2kb/+2kb from the TSS) and their overlap are shown. p
    Purification Gst Mnase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/purification gst mnase/product/Thermo Fisher
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    purification gst mnase - by Bioz Stars, 2020-05
    93/100 stars
      Buy from Supplier

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    DKC1-associated RNAs in recombinant DKC1 complexes are resistant to extensive MNase digestion. RNAs co-purified from mock and MNase-treated recombinant DKC1 complexes in Figure 4—figure supplement 1 are 5′ end radiolabeled and separated on a 6% urea-polyacrylamide gel as described in Figure 2—figure supplement 1 . Note that the prominently labeled 130–140 nucleotide (nt)-long RNA clusters are resistant to complete nuclease digestion. It appears that these RNAs are cut on average once by MNase to generate two new smaller clusters (80–90 nt and 30–55 nt) that remain stably associated with the DKC1 complexes. Increasing the amount of MNase and/or nuclease digestion time did not change the patterns or disrupt the integrity of the protein complexes (data not shown). DOI: http://dx.doi.org/10.7554/eLife.03573.010

    Journal: eLife

    Article Title: The dyskerin ribonucleoprotein complex as an OCT4/SOX2 coactivator in embryonic stem cells

    doi: 10.7554/eLife.03573

    Figure Lengend Snippet: DKC1-associated RNAs in recombinant DKC1 complexes are resistant to extensive MNase digestion. RNAs co-purified from mock and MNase-treated recombinant DKC1 complexes in Figure 4—figure supplement 1 are 5′ end radiolabeled and separated on a 6% urea-polyacrylamide gel as described in Figure 2—figure supplement 1 . Note that the prominently labeled 130–140 nucleotide (nt)-long RNA clusters are resistant to complete nuclease digestion. It appears that these RNAs are cut on average once by MNase to generate two new smaller clusters (80–90 nt and 30–55 nt) that remain stably associated with the DKC1 complexes. Increasing the amount of MNase and/or nuclease digestion time did not change the patterns or disrupt the integrity of the protein complexes (data not shown). DOI: http://dx.doi.org/10.7554/eLife.03573.010

    Article Snippet: Bound DKC1 complexes were treated with 300 U of MNase (Thermo Scientific, Waltham, MA) or buffer at room temperature and nutated for 1 hr.

    Techniques: Recombinant, Purification, Labeling, Stable Transfection

    Micrococcal nuclease (MNase)-treated recombinant DKC1 complexes remain structurally intact. Recombinant wild-type (WT) and various mutant DKC1 complexes are mock treated (−) or digested extensively with MNase (+), and washed extensively to remove any dissociated RNAs prior to FLAG peptide elution. Eluted protein complexes are analyzed by Coomassie Blue staining. DOI: http://dx.doi.org/10.7554/eLife.03573.009

    Journal: eLife

    Article Title: The dyskerin ribonucleoprotein complex as an OCT4/SOX2 coactivator in embryonic stem cells

    doi: 10.7554/eLife.03573

    Figure Lengend Snippet: Micrococcal nuclease (MNase)-treated recombinant DKC1 complexes remain structurally intact. Recombinant wild-type (WT) and various mutant DKC1 complexes are mock treated (−) or digested extensively with MNase (+), and washed extensively to remove any dissociated RNAs prior to FLAG peptide elution. Eluted protein complexes are analyzed by Coomassie Blue staining. DOI: http://dx.doi.org/10.7554/eLife.03573.009

    Article Snippet: Bound DKC1 complexes were treated with 300 U of MNase (Thermo Scientific, Waltham, MA) or buffer at room temperature and nutated for 1 hr.

    Techniques: Recombinant, Mutagenesis, Staining

    MNase digestion moderately increases DKC1 coactivator activity. Mock (−) or MNase-treated (+) WT and Nop10 R34W DKC1 complexes are assayed in in vitro transcription reactions (over a fourfold concentration range) supplemented with OCT4, SOX2, recombinant XPC complex and SCC-B. DOI: http://dx.doi.org/10.7554/eLife.03573.011

    Journal: eLife

    Article Title: The dyskerin ribonucleoprotein complex as an OCT4/SOX2 coactivator in embryonic stem cells

    doi: 10.7554/eLife.03573

    Figure Lengend Snippet: MNase digestion moderately increases DKC1 coactivator activity. Mock (−) or MNase-treated (+) WT and Nop10 R34W DKC1 complexes are assayed in in vitro transcription reactions (over a fourfold concentration range) supplemented with OCT4, SOX2, recombinant XPC complex and SCC-B. DOI: http://dx.doi.org/10.7554/eLife.03573.011

    Article Snippet: Bound DKC1 complexes were treated with 300 U of MNase (Thermo Scientific, Waltham, MA) or buffer at room temperature and nutated for 1 hr.

    Techniques: Activity Assay, In Vitro, Concentration Assay, Recombinant

    Release of nucleosomes and DAMPs from amino-acid-depleted HeLa cells. ( A ) An inverted microscopic image of HeLa cells in the condition of amino-acid depletion. Arrows designate dying HeLa cells. ( B ) Genomic sequences of glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ), Fas, cytochrome oxidase subunit 1 ( Co1 ) and ATP synthase subunit 6 ( ATP6 ) were PCR amplified from extracellularly released DNA, genomic or mitochondrial DNAs. ( C ) Inverted and fluorescent microscopic images were taken from amino-acid-deprived HeLa cells in the presence of SYTOX, a membrane-impermeable DNA dye. HeLa cells deprived of amino acids for 15 h were fluorescence stained with histone H1 or IL6 antibodies ( D ), histone H2A, H2B, H3 or H4 antibodies ( E ) or HMGB1, Hsp90 or ERp57 antibodies ( F ) in combination with histone H1 antibody and 4',6-diamidino-2-phenylindole (DAPI). ( G ) Amino-acid-deprived HeLa cells were stained with SYTOX to determine viability, fixed and stained with DAPI and histone H1 antibodies. ( H ) Amino-acid-deprived HeLa cells were untreated or treated with MNase (500 mU/ml) for 10 min. Released DNA was quantitated at the indicated times. Data from triplicate samples are presented as mean±S.D. ( I ) Conditioned media from amino-acid-deprived HeLa cells treated or untreated with MNase were western blotted with histone H1, 2B, H3, H4, IL6, ERp57, HMGB1 or Hsp90 antibodies. ( J ) Images captured every hour from live imaging of amino-acid-deprived HeLa cells with SYTOX (green) and DRAQ5, membrane-permeable DNA dye (red). ( K ) SYTOX fluorescent intensities were measured from circularized areas of live imaging of amino-acid-deprived cells in 5-min intervals. ( L ) TEM images of control cells ( L a) and amino-acid-deprived HeLa cells ( L b– L d). ( M ) Amino-acid-deprived HeLa cells were fluorescence stained with lamin and nuclear pore antibodies, or lamin antibody and wheat germ agglutinin (WGE)

    Journal: Cell Death & Disease

    Article Title: Caspase-dependent cell death-associated release of nucleosome and damage-associated molecular patterns

    doi: 10.1038/cddis.2014.450

    Figure Lengend Snippet: Release of nucleosomes and DAMPs from amino-acid-depleted HeLa cells. ( A ) An inverted microscopic image of HeLa cells in the condition of amino-acid depletion. Arrows designate dying HeLa cells. ( B ) Genomic sequences of glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ), Fas, cytochrome oxidase subunit 1 ( Co1 ) and ATP synthase subunit 6 ( ATP6 ) were PCR amplified from extracellularly released DNA, genomic or mitochondrial DNAs. ( C ) Inverted and fluorescent microscopic images were taken from amino-acid-deprived HeLa cells in the presence of SYTOX, a membrane-impermeable DNA dye. HeLa cells deprived of amino acids for 15 h were fluorescence stained with histone H1 or IL6 antibodies ( D ), histone H2A, H2B, H3 or H4 antibodies ( E ) or HMGB1, Hsp90 or ERp57 antibodies ( F ) in combination with histone H1 antibody and 4',6-diamidino-2-phenylindole (DAPI). ( G ) Amino-acid-deprived HeLa cells were stained with SYTOX to determine viability, fixed and stained with DAPI and histone H1 antibodies. ( H ) Amino-acid-deprived HeLa cells were untreated or treated with MNase (500 mU/ml) for 10 min. Released DNA was quantitated at the indicated times. Data from triplicate samples are presented as mean±S.D. ( I ) Conditioned media from amino-acid-deprived HeLa cells treated or untreated with MNase were western blotted with histone H1, 2B, H3, H4, IL6, ERp57, HMGB1 or Hsp90 antibodies. ( J ) Images captured every hour from live imaging of amino-acid-deprived HeLa cells with SYTOX (green) and DRAQ5, membrane-permeable DNA dye (red). ( K ) SYTOX fluorescent intensities were measured from circularized areas of live imaging of amino-acid-deprived cells in 5-min intervals. ( L ) TEM images of control cells ( L a) and amino-acid-deprived HeLa cells ( L b– L d). ( M ) Amino-acid-deprived HeLa cells were fluorescence stained with lamin and nuclear pore antibodies, or lamin antibody and wheat germ agglutinin (WGE)

    Article Snippet: Released DNA from dying cells were digested with 500 mU/ml MNase (Thermo Scientific) for 5 min. Nuclease activity was stopped with 5 mM EDTA and the culture supernatants were collected and stored at −20 °C until quantification.

    Techniques: Genomic Sequencing, Polymerase Chain Reaction, Amplification, Fluorescence, Staining, Western Blot, Imaging, Transmission Electron Microscopy

    MapR and RHΔ CUT RUN Signals Are Enriched at Similar Regions Genome-wide (A) Schematic of RHΔC R using FLAG M2 antibody (left) and MapR using GST-RHΔ-MNase (right) in HEK293. (B) Enriched regions identified by RHΔC R and R-ChIP in HEK293. GRO-seq and H3K4me3 tracks indicate active gene transcription. (C) Venn diagram of gene-level overlap between RHΔC R and R-ChIP. Total number of unique genes with an R-loop at the promoter region (−2kb/+2kb from the TSS) and their overlap are shown. p

    Journal: Cell reports

    Article Title: Mapping Native R-Loops Genome-wide Using a Targeted Nuclease Approach

    doi: 10.1016/j.celrep.2019.09.052

    Figure Lengend Snippet: MapR and RHΔ CUT RUN Signals Are Enriched at Similar Regions Genome-wide (A) Schematic of RHΔC R using FLAG M2 antibody (left) and MapR using GST-RHΔ-MNase (right) in HEK293. (B) Enriched regions identified by RHΔC R and R-ChIP in HEK293. GRO-seq and H3K4me3 tracks indicate active gene transcription. (C) Venn diagram of gene-level overlap between RHΔC R and R-ChIP. Total number of unique genes with an R-loop at the promoter region (−2kb/+2kb from the TSS) and their overlap are shown. p

    Article Snippet: Protein Expression and Purification GST-MNase and GST-RHΔMNase were cloned into pGEX plasmid and transformed into BL21 (DE3) (ThermoFisher C601003) for expression.

    Techniques: Genome Wide, Chromatin Immunoprecipitation

    MapR, a Native and Antibody-Independent R-Loop Detection Strategy R-loop recognition and recovery by MapR. Step 1: cells are immobilized on concanavalin A beads and permeabilized. Step 2: equimolar amounts of a catalytic deficient mutant of RNase H fused to micrococcal nuclease (GST-RHΔ-MNase) or GST-MNase is added to immobilized cells. Step 3: the RHΔ module recognizes and binds R-loops on chromatin. Step 4: controlled activation of the MNase moiety by addition of calcium results in cleavage of DNA fragments in proximity to R-loops. Step 5: Released R-loops diffuse out of the cell; the DNA is recovered and sequenced.

    Journal: Cell reports

    Article Title: Mapping Native R-Loops Genome-wide Using a Targeted Nuclease Approach

    doi: 10.1016/j.celrep.2019.09.052

    Figure Lengend Snippet: MapR, a Native and Antibody-Independent R-Loop Detection Strategy R-loop recognition and recovery by MapR. Step 1: cells are immobilized on concanavalin A beads and permeabilized. Step 2: equimolar amounts of a catalytic deficient mutant of RNase H fused to micrococcal nuclease (GST-RHΔ-MNase) or GST-MNase is added to immobilized cells. Step 3: the RHΔ module recognizes and binds R-loops on chromatin. Step 4: controlled activation of the MNase moiety by addition of calcium results in cleavage of DNA fragments in proximity to R-loops. Step 5: Released R-loops diffuse out of the cell; the DNA is recovered and sequenced.

    Article Snippet: Protein Expression and Purification GST-MNase and GST-RHΔMNase were cloned into pGEX plasmid and transformed into BL21 (DE3) (ThermoFisher C601003) for expression.

    Techniques: Mutagenesis, Activation Assay