exonuclease deficient dna polymerase  (New England Biolabs)


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    Name:
    DNA Polymerase I Klenow Fragment
    Description:
    DNA Polymerase I Klenow Fragment 1 000 units
    Catalog Number:
    M0210L
    Price:
    248
    Category:
    DNA Polymerases
    Size:
    1 000 units
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    Structured Review

    New England Biolabs exonuclease deficient dna polymerase
    DNA Polymerase I Klenow Fragment
    DNA Polymerase I Klenow Fragment 1 000 units
    https://www.bioz.com/result/exonuclease deficient dna polymerase/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    exonuclease deficient dna polymerase - by Bioz Stars, 2021-07
    99/100 stars

    Images

    1) Product Images from "Single-stranded nucleic acid sensing and coacervation by linker histone H1"

    Article Title: Single-stranded nucleic acid sensing and coacervation by linker histone H1

    Journal: bioRxiv

    doi: 10.1101/2021.03.17.435841

    H1 coalesces around nascent ssDNA. ( A ) Schematic of the combined single-molecule fluorescence and force microscopy. A biotinylated λ-DNA molecule (48.5 kbp) is tethered between two streptavidin-coated polystyrene beads. ( B ) A representative kymograph of Cy3-H1 binding to DNA over time as the inter-bead distance was increased. ( C ) Total H1 signal across the DNA as a function of time for the kymograph shown in (B). ( D ) Distribution of the H1 signal along the DNA at two specific time points (T1 and T2) as indicated by the arrows in (B). ( E ) Cartoon illustrating the distinct binding configurations of H1 on DNA under different tensions. ssDNA is created by force-induced unpeeling. ( F ) Schematic of two-color imaging for simultaneous visualization of H1 and RPA binding to DNA. ( G ) A representative kymograph of Cy3-H1 (green) and AlexaFluor488-RPA (blue) binding to DNA over time as the inter-bead distance was increased. ( H ) Total H1 and RPA signals across the DNA as a function of time for the kymograph shown in (G). ( I ) Distribution of the H1 (green) and RPA (blue) signals along the DNA at a specific time point (T1) as indicated by the arrow in (G). ( J ) Cartoon illustrating that H1 and RPA occupy separate regions of the tethered DNA. H1 coalesces around relaxed ssDNA, whereas RPA binds to ssDNA under tension.
    Figure Legend Snippet: H1 coalesces around nascent ssDNA. ( A ) Schematic of the combined single-molecule fluorescence and force microscopy. A biotinylated λ-DNA molecule (48.5 kbp) is tethered between two streptavidin-coated polystyrene beads. ( B ) A representative kymograph of Cy3-H1 binding to DNA over time as the inter-bead distance was increased. ( C ) Total H1 signal across the DNA as a function of time for the kymograph shown in (B). ( D ) Distribution of the H1 signal along the DNA at two specific time points (T1 and T2) as indicated by the arrows in (B). ( E ) Cartoon illustrating the distinct binding configurations of H1 on DNA under different tensions. ssDNA is created by force-induced unpeeling. ( F ) Schematic of two-color imaging for simultaneous visualization of H1 and RPA binding to DNA. ( G ) A representative kymograph of Cy3-H1 (green) and AlexaFluor488-RPA (blue) binding to DNA over time as the inter-bead distance was increased. ( H ) Total H1 and RPA signals across the DNA as a function of time for the kymograph shown in (G). ( I ) Distribution of the H1 (green) and RPA (blue) signals along the DNA at a specific time point (T1) as indicated by the arrow in (G). ( J ) Cartoon illustrating that H1 and RPA occupy separate regions of the tethered DNA. H1 coalesces around relaxed ssDNA, whereas RPA binds to ssDNA under tension.

    Techniques Used: Fluorescence, Microscopy, Binding Assay, Imaging, Recombinase Polymerase Amplification

    2) Product Images from "Visual Detection of Gene Mutations Based on Isothermal Strand-Displacement Polymerase Reaction and Lateral Flow Strip"

    Article Title: Visual Detection of Gene Mutations Based on Isothermal Strand-Displacement Polymerase Reaction and Lateral Flow Strip

    Journal: Biosensors & bioelectronics

    doi: 10.1016/j.bios.2011.10.037

    (A) The optical responses of 1-nM R156H-mutant DNA on the LFSs with different ISDPR solutions. Solution 1: 5.0×10 −8 M biotin-modified hairpin probe, 5.0×10 −8 M digoxin-modified primer, 3-U polymerase Klenow fragment exo
    Figure Legend Snippet: (A) The optical responses of 1-nM R156H-mutant DNA on the LFSs with different ISDPR solutions. Solution 1: 5.0×10 −8 M biotin-modified hairpin probe, 5.0×10 −8 M digoxin-modified primer, 3-U polymerase Klenow fragment exo

    Techniques Used: Mutagenesis, Modification

    3) Product Images from "Sensitive fluorescence detection of nucleic acids based on isothermal circular strand-displacement polymerization reaction"

    Article Title: Sensitive fluorescence detection of nucleic acids based on isothermal circular strand-displacement polymerization reaction

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkn1024

    Detection of different concentrations of target based on isothermal strand-displacement polymerization reaction. Experiments were performed in the presence of 15U polymerase Klenow fragment exo − and 100 μM dNTPs with 5 × 10 –8 M probe, 5 × 10 –8 M primer, and different concentrations of target. ( A ) Monitoring the fluorescence intensity of this amplified DNA detection method over a range of target DNA concentrations. The curves from a to i contain the target with 1.0 × 10 –10 , 2.0 × 10 –11 , 4.0 × 10 –12 , 8.0 × 10 –13 , 1.6 × 10 –13 , 3.2 × 10 –14 , 6.4 × 10 –15 , 1.28 × 10 –15 and 0 M, respectively. All samples were incubated at 37°C. ( B ) The relationship of the rate of fluorescence enhancement with target DNA concentration.
    Figure Legend Snippet: Detection of different concentrations of target based on isothermal strand-displacement polymerization reaction. Experiments were performed in the presence of 15U polymerase Klenow fragment exo − and 100 μM dNTPs with 5 × 10 –8 M probe, 5 × 10 –8 M primer, and different concentrations of target. ( A ) Monitoring the fluorescence intensity of this amplified DNA detection method over a range of target DNA concentrations. The curves from a to i contain the target with 1.0 × 10 –10 , 2.0 × 10 –11 , 4.0 × 10 –12 , 8.0 × 10 –13 , 1.6 × 10 –13 , 3.2 × 10 –14 , 6.4 × 10 –15 , 1.28 × 10 –15 and 0 M, respectively. All samples were incubated at 37°C. ( B ) The relationship of the rate of fluorescence enhancement with target DNA concentration.

    Techniques Used: Fluorescence, Amplification, Incubation, Concentration Assay

    Related Articles

    other:

    Article Title: An Efficient Strategy for Broad-Range Detection of Low Abundance Bacteria without DNA Decontamination of PCR Reagents
    Article Snippet: MaterialsThe exo I and Klenow DNA polymerase were purchased from New England Biolab (Ipswich, MA).

    Purification:

    Article Title: Zinc-finger nuclease-driven targeted integration into mammalian genomes using donors with limited chromosomal homology
    Article Snippet: Reactions with AAVS1 , IL2Rγ and HindIII were terminated after 2 min and the GS reaction after 5 min by addition of 10 mM Tris/1 mM EDTA to 200 µl, followed by phenol extraction and ethanol precipitation. .. Linearized plasmids were gel purified by agarose gel electrophoresis and incubated for 30 min at 37°C with 0.05 U Klenow DNA polymerase (New England Biolabs) in 1 × Buffer 2, plus 50 µM dNTPs. .. Klenow polymerase was inactivated by incubation at 75°C for 20 min, followed by addition of 20 U of T4 DNA Ligase (New England Biolabs) and ATP to 1 mM.

    Agarose Gel Electrophoresis:

    Article Title: Zinc-finger nuclease-driven targeted integration into mammalian genomes using donors with limited chromosomal homology
    Article Snippet: Reactions with AAVS1 , IL2Rγ and HindIII were terminated after 2 min and the GS reaction after 5 min by addition of 10 mM Tris/1 mM EDTA to 200 µl, followed by phenol extraction and ethanol precipitation. .. Linearized plasmids were gel purified by agarose gel electrophoresis and incubated for 30 min at 37°C with 0.05 U Klenow DNA polymerase (New England Biolabs) in 1 × Buffer 2, plus 50 µM dNTPs. .. Klenow polymerase was inactivated by incubation at 75°C for 20 min, followed by addition of 20 U of T4 DNA Ligase (New England Biolabs) and ATP to 1 mM.

    Incubation:

    Article Title: Zinc-finger nuclease-driven targeted integration into mammalian genomes using donors with limited chromosomal homology
    Article Snippet: Reactions with AAVS1 , IL2Rγ and HindIII were terminated after 2 min and the GS reaction after 5 min by addition of 10 mM Tris/1 mM EDTA to 200 µl, followed by phenol extraction and ethanol precipitation. .. Linearized plasmids were gel purified by agarose gel electrophoresis and incubated for 30 min at 37°C with 0.05 U Klenow DNA polymerase (New England Biolabs) in 1 × Buffer 2, plus 50 µM dNTPs. .. Klenow polymerase was inactivated by incubation at 75°C for 20 min, followed by addition of 20 U of T4 DNA Ligase (New England Biolabs) and ATP to 1 mM.

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  • 99
    New England Biolabs t7 endonuclease i
    Deletion of MCM10 through CRISPR/Cas9 technology. Notes:  ( A ) Validation of CRISPR/Cas9-mediated knockout efficiency using a mCherry/GFP reporter construct. EC109 cells were transfected with the Cas9, sgRNAs, and reporter constructs, and mCherry/GFP fluorescence was examined 48 h after transfection. (a) Bright field image of cells. (b) Some cells displayed GFP fluorescence, indicating the presence of CRISPR/Cas9-mediated removal of target sequence. (c) EC109 cells that were transfected with reporter construct showed mCherry fluorescence. (d) Merged image of green and red fluorescence yielded yellow fluorescence. Scale bar = 100 µm. ( B ) T7 endonuclease assay. Different clones derived from EC109 cells transfected with Cas9 and sgRNAs were subjected to PCR amplification of genomic DNA containing sgRNA-1 target site. The size of T7 endonuclease I-digested DNA fragments is indicated on the right. Control, negative control. ( C ) Upper; RT-PCR analysis of MCM10 mRNA expression in different EC109 sublines. Lower; Western blot analysis of MCM10 protein levels. ( D ) Depletion of MCM10 hampers the migration of ESCC cells. In vitro wound-healing assay was performed to assess cell migration capacity. Top; one representative experiment. The percentage of wound closure was determined from three independent experiments. * P
    T7 Endonuclease I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs klenow fragment exo buffer
    Fluorescence intensity of the proposed aptamer sensing with isothermal circular system containing the DNA aptamer (1 μM), signaling probe (1 μM), primer (2.5 μM), dNTPs (2 mM), <t>Klenow</t> Fragment <t>exo-</t> (1 U/μL), and Nt.BbvCI (1 U/μL), and initiated with a different amount of the PDGF-BB sample (a: 0, b: 0.1 ng/mL, c: 1 ng/mL, d: 10 f ng/mL, e: 20 ng/mL, f: 40 ng/mL, g: 60 ng/mL) at different times.
    Klenow Fragment Exo Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs dna polymerase i
    PAGE analysis of primer extension experiments with single OXP-modified and PDE primers. Primer extension with Klenow fragment of <t>DNA</t> <t>polymerase</t> I of nonheated ( A ) and preheated ( B ) single OXP-modified reverse primer, respectively along template 2. The extension reactions were incubated at 25°C for the indicated times after which the reaction mixtures were quenched and analyzed. ( C ) Primer extension with Taq DNA polymerase of PDE and OXP forward primers (nonheated control and preheated sample) along template oligonucleotide 1. Extension reactions were incubated at 25°C for 15 min, after which the aliquots from reaction mixtures were quenched and analyzed.
    Dna Polymerase I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs dna polymerase i klenow fragment
    Inhibition of Pol I results in <t>DNA</t> damage in a subset of cells a , Representative immunofluorescence images of wild-type and TCOF1 +/− cNCCs stained with an antibody against γH2A.X; quantification is shown in b . c , Representative immunofluorescence images of DNA-damaged wild-type cNCCs stained with an antibody against γH2A.X after 1 h treatment with iPol I or actinomycin D (ActD); quantification is shown in d . e , Representative immunofluorescence images of DNA-damaged HeLa cells stained with an antibody against γH2A.X after 1 h treatment with iPol I; quantification is shown in f . For a – f , cells were collected from n = 3 biologically independent experiments. Boxes represent median value and 25th and 75th percentiles, whiskers are minimum to maximum, crosses are outliers. ***P
    Dna Polymerase I Klenow Fragment, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Deletion of MCM10 through CRISPR/Cas9 technology. Notes:  ( A ) Validation of CRISPR/Cas9-mediated knockout efficiency using a mCherry/GFP reporter construct. EC109 cells were transfected with the Cas9, sgRNAs, and reporter constructs, and mCherry/GFP fluorescence was examined 48 h after transfection. (a) Bright field image of cells. (b) Some cells displayed GFP fluorescence, indicating the presence of CRISPR/Cas9-mediated removal of target sequence. (c) EC109 cells that were transfected with reporter construct showed mCherry fluorescence. (d) Merged image of green and red fluorescence yielded yellow fluorescence. Scale bar = 100 µm. ( B ) T7 endonuclease assay. Different clones derived from EC109 cells transfected with Cas9 and sgRNAs were subjected to PCR amplification of genomic DNA containing sgRNA-1 target site. The size of T7 endonuclease I-digested DNA fragments is indicated on the right. Control, negative control. ( C ) Upper; RT-PCR analysis of MCM10 mRNA expression in different EC109 sublines. Lower; Western blot analysis of MCM10 protein levels. ( D ) Depletion of MCM10 hampers the migration of ESCC cells. In vitro wound-healing assay was performed to assess cell migration capacity. Top; one representative experiment. The percentage of wound closure was determined from three independent experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Ablation of MCM10 using CRISPR/Cas9 restrains the growth and migration of esophageal squamous cell carcinoma cells through inhibition of Akt signaling

    doi: 10.2147/OTT.S157025

    Figure Lengend Snippet: Deletion of MCM10 through CRISPR/Cas9 technology. Notes: ( A ) Validation of CRISPR/Cas9-mediated knockout efficiency using a mCherry/GFP reporter construct. EC109 cells were transfected with the Cas9, sgRNAs, and reporter constructs, and mCherry/GFP fluorescence was examined 48 h after transfection. (a) Bright field image of cells. (b) Some cells displayed GFP fluorescence, indicating the presence of CRISPR/Cas9-mediated removal of target sequence. (c) EC109 cells that were transfected with reporter construct showed mCherry fluorescence. (d) Merged image of green and red fluorescence yielded yellow fluorescence. Scale bar = 100 µm. ( B ) T7 endonuclease assay. Different clones derived from EC109 cells transfected with Cas9 and sgRNAs were subjected to PCR amplification of genomic DNA containing sgRNA-1 target site. The size of T7 endonuclease I-digested DNA fragments is indicated on the right. Control, negative control. ( C ) Upper; RT-PCR analysis of MCM10 mRNA expression in different EC109 sublines. Lower; Western blot analysis of MCM10 protein levels. ( D ) Depletion of MCM10 hampers the migration of ESCC cells. In vitro wound-healing assay was performed to assess cell migration capacity. Top; one representative experiment. The percentage of wound closure was determined from three independent experiments. * P

    Article Snippet: After treatment with T7 endonuclease I (New England Biolabs, Ipswich, MA, USA) at 37°C for 2 h, the resulting fragments were subjected to 1% agarose gel electrophoresis and stained with ethidium bromide.

    Techniques: CRISPR, Knock-Out, Construct, Transfection, Fluorescence, Sequencing, Clone Assay, Derivative Assay, Polymerase Chain Reaction, Amplification, Negative Control, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Migration, In Vitro, Wound Healing Assay

    Fluorescence intensity of the proposed aptamer sensing with isothermal circular system containing the DNA aptamer (1 μM), signaling probe (1 μM), primer (2.5 μM), dNTPs (2 mM), Klenow Fragment exo- (1 U/μL), and Nt.BbvCI (1 U/μL), and initiated with a different amount of the PDGF-BB sample (a: 0, b: 0.1 ng/mL, c: 1 ng/mL, d: 10 f ng/mL, e: 20 ng/mL, f: 40 ng/mL, g: 60 ng/mL) at different times.

    Journal: Sensors (Basel, Switzerland)

    Article Title: Aptamer Conformation Switching-Induced Two-Stage Amplification for Fluorescent Detection of Proteins

    doi: 10.3390/s19010077

    Figure Lengend Snippet: Fluorescence intensity of the proposed aptamer sensing with isothermal circular system containing the DNA aptamer (1 μM), signaling probe (1 μM), primer (2.5 μM), dNTPs (2 mM), Klenow Fragment exo- (1 U/μL), and Nt.BbvCI (1 U/μL), and initiated with a different amount of the PDGF-BB sample (a: 0, b: 0.1 ng/mL, c: 1 ng/mL, d: 10 f ng/mL, e: 20 ng/mL, f: 40 ng/mL, g: 60 ng/mL) at different times.

    Article Snippet: The deoxynucleotide solution mixture (dNTPs), polymerase Klenow Fragment exo- (10 U/μL) accompanied by 10× Klenow Fragment exo- buffer, and the nicking endonuclease Nt.BbvCI accompanied by 10× New England Biolabs (NEB) buffer were purchased from New England Biolabs Ltd (Beijing, China).

    Techniques: Fluorescence

    The fluorescence spectrum of the developed sensing system is collected in a blank control sample (curve “a”); in the presence of aptamer DNA, an MB, polymerase, and nicking endonuclease, but without PDGF-BB (curve “b”); in the presence of PDGF-BB, aptamer DNA, and an MB (curve “c”); in the presence of PDGF-BB, aptamer DNA, an MB, and polymerase (curve “d”); and in the presence of PDGF-BB, aptamer DNA, MB, polymerase, and nicking endonuclease Nt.BbvCI (curve “e”). The reaction is in an NEB buffer (pH 7.9) at 37 °C for 2 h containing 1 μM aptamer DNA, 1 μM MB, 2.5 μM primer, 100 ng/mL PDGF-BB, 1 U/μL Klenow Fragment exo-, 2 mM dNTPs, and 1 U/μL Nt.BbvCI.

    Journal: Sensors (Basel, Switzerland)

    Article Title: Aptamer Conformation Switching-Induced Two-Stage Amplification for Fluorescent Detection of Proteins

    doi: 10.3390/s19010077

    Figure Lengend Snippet: The fluorescence spectrum of the developed sensing system is collected in a blank control sample (curve “a”); in the presence of aptamer DNA, an MB, polymerase, and nicking endonuclease, but without PDGF-BB (curve “b”); in the presence of PDGF-BB, aptamer DNA, and an MB (curve “c”); in the presence of PDGF-BB, aptamer DNA, an MB, and polymerase (curve “d”); and in the presence of PDGF-BB, aptamer DNA, MB, polymerase, and nicking endonuclease Nt.BbvCI (curve “e”). The reaction is in an NEB buffer (pH 7.9) at 37 °C for 2 h containing 1 μM aptamer DNA, 1 μM MB, 2.5 μM primer, 100 ng/mL PDGF-BB, 1 U/μL Klenow Fragment exo-, 2 mM dNTPs, and 1 U/μL Nt.BbvCI.

    Article Snippet: The deoxynucleotide solution mixture (dNTPs), polymerase Klenow Fragment exo- (10 U/μL) accompanied by 10× Klenow Fragment exo- buffer, and the nicking endonuclease Nt.BbvCI accompanied by 10× New England Biolabs (NEB) buffer were purchased from New England Biolabs Ltd (Beijing, China).

    Techniques: Fluorescence

    PAGE analysis of primer extension experiments with single OXP-modified and PDE primers. Primer extension with Klenow fragment of DNA polymerase I of nonheated ( A ) and preheated ( B ) single OXP-modified reverse primer, respectively along template 2. The extension reactions were incubated at 25°C for the indicated times after which the reaction mixtures were quenched and analyzed. ( C ) Primer extension with Taq DNA polymerase of PDE and OXP forward primers (nonheated control and preheated sample) along template oligonucleotide 1. Extension reactions were incubated at 25°C for 15 min, after which the aliquots from reaction mixtures were quenched and analyzed.

    Journal: Nucleic Acids Research

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance

    doi: 10.1093/nar/gkn575

    Figure Lengend Snippet: PAGE analysis of primer extension experiments with single OXP-modified and PDE primers. Primer extension with Klenow fragment of DNA polymerase I of nonheated ( A ) and preheated ( B ) single OXP-modified reverse primer, respectively along template 2. The extension reactions were incubated at 25°C for the indicated times after which the reaction mixtures were quenched and analyzed. ( C ) Primer extension with Taq DNA polymerase of PDE and OXP forward primers (nonheated control and preheated sample) along template oligonucleotide 1. Extension reactions were incubated at 25°C for 15 min, after which the aliquots from reaction mixtures were quenched and analyzed.

    Article Snippet: Primer extension with Klenow fragment of DNA polymerase Primer extension experiments using large fragment (Klenow) of DNA polymerase I (New England Biolabs) were performed at 25°C using the HIV-1 tat reverse primer (5′-AATACTATGGTCCACACAACTATTGCT-3′) that was unmodified or contained a single OXP modification.

    Techniques: Polyacrylamide Gel Electrophoresis, Modification, Incubation

    Inhibition of Pol I results in DNA damage in a subset of cells a , Representative immunofluorescence images of wild-type and TCOF1 +/− cNCCs stained with an antibody against γH2A.X; quantification is shown in b . c , Representative immunofluorescence images of DNA-damaged wild-type cNCCs stained with an antibody against γH2A.X after 1 h treatment with iPol I or actinomycin D (ActD); quantification is shown in d . e , Representative immunofluorescence images of DNA-damaged HeLa cells stained with an antibody against γH2A.X after 1 h treatment with iPol I; quantification is shown in f . For a – f , cells were collected from n = 3 biologically independent experiments. Boxes represent median value and 25th and 75th percentiles, whiskers are minimum to maximum, crosses are outliers. ***P

    Journal: Nature

    Article Title: Tissue–selective effects of nucleolar stress and rDNA damage in developmental disorders

    doi: 10.1038/nature25449

    Figure Lengend Snippet: Inhibition of Pol I results in DNA damage in a subset of cells a , Representative immunofluorescence images of wild-type and TCOF1 +/− cNCCs stained with an antibody against γH2A.X; quantification is shown in b . c , Representative immunofluorescence images of DNA-damaged wild-type cNCCs stained with an antibody against γH2A.X after 1 h treatment with iPol I or actinomycin D (ActD); quantification is shown in d . e , Representative immunofluorescence images of DNA-damaged HeLa cells stained with an antibody against γH2A.X after 1 h treatment with iPol I; quantification is shown in f . For a – f , cells were collected from n = 3 biologically independent experiments. Boxes represent median value and 25th and 75th percentiles, whiskers are minimum to maximum, crosses are outliers. ***P

    Article Snippet: After the NT2 wash, DDX21-bound RNA–protein complexes were dephosphorylated with T4 PNK (NEB, catalogue number M0210) for 30 min in an Eppendorf Thermomixer at 37 °C, 15 s at 1,400 r.p.m., 90 s rest in a 30 μl reaction, pH 6.5, containing 10 units of T4 PNK, 0.1 μl SUPERase-IN, and 6 μl of PEG-400 (16.7% final).

    Techniques: Inhibition, Immunofluorescence, Staining