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Roche dnaase i
Dnaase I, supplied by Roche, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/dnaase i/product/Roche
Average 95 stars, based on 2 article reviews
Price from $9.99 to $1999.99
dnaase i - by Bioz Stars, 2020-04
95/100 stars

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Related Articles

Centrifugation:

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: The nuclear lysates were diluted with 2 volumes of 10 m m HEPES and cleared by centrifugation at 13,000 × g . .. For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added.

Article Title: Characterization of the Single Stranded DNA Binding Protein SsbB Encoded in the Gonoccocal Genetic Island
Article Snippet: After 3 hrs the cells were harvested by centrifugation, resuspended in 30 ml of buffer A (50 mM NaPO4 , 300 mM NaCl, pH 8.0) and stored at −80°C. .. After thawing, the solution was supplemented with 1 tablet of Protease Inhibitor Cocktail (Roche) and 1 mgr of DNase I (Roche), and the cells were disrupted 3 times in a high-pressure Cell Disrupter (Constant Cell Disruption Systems) at 2.300 bar.

Stable Transfection:

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: For CoIP of NFAT-containing complexes or control proteins, 2–3·108 stably transfected Jurkat cells were stimulated for 2 h with PMA/ionomycin. .. For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added.

Construct:

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: The fusion constructs of PaExoY and VnExoY with an N-terminal maltose-binding protein (MBP, 40.4 kDa), designed as follows: (His-Tag)-(MBP)-(PreScission-site)-(PaExoY/VnExoY)-(Strep-tagII) and referred in the text as MBP-PaExoY or MBP-VnExoY were purified under non-denaturing conditions successively from HisTrap, StrepTrap, and Superdex 200 16/60 columns using standard protocols. .. Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ].

Electrophoresis:

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added. .. To monitor DNA digestion, DNA was extracted from a portion of the extracts using spin column purification (Macherey Nagel) and separated by electrophoresis on a 1% GelRed-agarose gel.

Incubation:

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: The suspension was spun for 1 min at 8000 × g, and the pellet (containing nuclei) was resuspended in high salt buffer (500 m m NaCl, 25 m m HEPES, 2 m m EDTA, 0.5% Tween 20), incubated on ice for 30 min, subjected to ultrasound shearing, and incubated again on ice for 30 min. .. For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added.

Article Title: Dead or alive: Deoxyribonuclease I sensitive bacteria and implications for the sinus microbiome
Article Snippet: Duplicate samples were placed in separate, sterile tubes and immerged in sufficient diluted DNase I buffer per manufacture standard protocol (Roche, Basel, Switzerland); 10 U/μL DNase I (Roche) was added to one sample in each pair. .. The samples were incubated at 37°C for 10 minutes to allow for DNase I-mediated degradation of free DNA, followed by 75°C for 10 minutes to deactivate the DNase I.

Activity Assay:

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: Activity assays to verify the functionality of the MBP-fusion proteins showed similar specific activities as the corresponding PaExoY or VnExoY enzymes for the synthesis of cGMP or cAMP, respectively. .. Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ].

Expressing:

Article Title: Characterization of the Single Stranded DNA Binding Protein SsbB Encoded in the Gonoccocal Genetic Island
Article Snippet: Paragraph title: Expression and purification of SsbB ... After thawing, the solution was supplemented with 1 tablet of Protease Inhibitor Cocktail (Roche) and 1 mgr of DNase I (Roche), and the cells were disrupted 3 times in a high-pressure Cell Disrupter (Constant Cell Disruption Systems) at 2.300 bar.

Derivative Assay:

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: .. Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ]. .. D . melanogaster cytoplasmic wild-type (Dm-actin, UNIPROT accession number: ACT1_DROME) and mutant actin (actin NP) were produced and purified from SF9 insect cells infected with recombinant Baculoviruses as described before [ ].

Transfection:

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: For CoIP of NFAT-containing complexes or control proteins, 2–3·108 stably transfected Jurkat cells were stimulated for 2 h with PMA/ionomycin. .. For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added.

Protease Inhibitor:

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added. .. All buffers were supplemented with complete protease inhibitor (Roche Applied Science), phosphatase inhibitors (2.5 m m sodium pyrophosphate, 1 m m 2-glycerol phosphate, 1 m m sodium orthovanadate), and NFAT kinase inhibitors (2 μ m JNK inhibitor VII, 2 μ m kenpaullone, 2 μ m PD169-316, 10 μ m ellagic acid).

Article Title: Characterization of the Single Stranded DNA Binding Protein SsbB Encoded in the Gonoccocal Genetic Island
Article Snippet: .. After thawing, the solution was supplemented with 1 tablet of Protease Inhibitor Cocktail (Roche) and 1 mgr of DNase I (Roche), and the cells were disrupted 3 times in a high-pressure Cell Disrupter (Constant Cell Disruption Systems) at 2.300 bar. .. Cell debris was removed by centrifugation at 10.000 rpm in an F10-6x500y rotor (FiberLite) and the supernatant was filtered through a 0.45 µm filter.

Infection:

Article Title: Discordant Results Obtained with Francisella tularensis during In Vitro and In Vivo Immunological Studies Are Attributable to Compromised Bacterial Structural Integrity
Article Snippet: .. ***P < 0.001. (All results shown were subjected to One-way ANOVA with Bonferroni's Post-test). (E) & (F) Wild-type and AIM2−/− BMDMs were infected with Ft LVS cultivated in MHB either untreated or treated with DNase I at a MOI of 100. .. Supernatants collected after 24 h were analyzed for the presence of cytokines by CBA.

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ]. .. D . melanogaster cytoplasmic wild-type (Dm-actin, UNIPROT accession number: ACT1_DROME) and mutant actin (actin NP) were produced and purified from SF9 insect cells infected with recombinant Baculoviruses as described before [ ].

other:

Article Title: Transcriptional regulation of BRCA1 expression by a metabolic switch
Article Snippet: For each DNase I digestion, approximately 1×106 nuclei were harvested and resuspended in 200 ul of pre-warmed (37 ° C) Buffer A, supplemented with 6 mM CaCl2 , 75 mM NaCl, and the DNase I (0, 170, 340, and 680 units).

Article Title: A refined technique for extraction of extracellular matrices from bacterial biofilms and its applicability
Article Snippet: ECM-degrading enzymes Dispersin B from Aggregatibacter actinomycetemcomitans (20 μg ml−1 , Kane Biotech Inc., Manitoba, Canada), Proteinase K from Tritirachium album (100 μg ml−1 , Sigma, St Louis, MO, USA) and DNase I (100 U ml−1 , Roche Diagnostics, Mannheim, Germany) were used for dispersal of preformed biofilms or degradation of ECM components.

Article Title: Extracellular DNA facilitates bacterial adhesion during Burkholderia pseudomallei biofilm formation
Article Snippet: DNase I noticeably lowered eDNA concentrations in biofilm if the enzyme was added into the starting inoculum (0 h).

Recombinant:

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ]. .. D . melanogaster cytoplasmic wild-type (Dm-actin, UNIPROT accession number: ACT1_DROME) and mutant actin (actin NP) were produced and purified from SF9 insect cells infected with recombinant Baculoviruses as described before [ ].

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: .. For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added. ..

Animal Model:

Article Title: Neutrophil Extracellular Traps Are Pathogenic in Ventilator-Induced Lung Injury and Partially Dependent on TLR4
Article Snippet: .. Discussion In this study, we illustrated the following: ventilation with high-tidal volume causes lung injury and inflammation in mice; NETs increase in the lung of the VILI animal model upon the elevation of known NET-associated proteins and extracellular DNA; pretreatment of NET degradation with DNase I alleviates the preceding lung injury, which means that NET formation has an adverse effect on the development of VILI; and TLR4 is involved in NET formation in VILI in mice. ..

Mutagenesis:

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ]. .. D . melanogaster cytoplasmic wild-type (Dm-actin, UNIPROT accession number: ACT1_DROME) and mutant actin (actin NP) were produced and purified from SF9 insect cells infected with recombinant Baculoviruses as described before [ ].

Co-Immunoprecipitation Assay:

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: Lysis protocol was adopted from Klenova et al. ( ) and was used for all CoIP experiments, including the CoIP-MS experiments. .. For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added.

Purification:

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: .. Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ]. .. D . melanogaster cytoplasmic wild-type (Dm-actin, UNIPROT accession number: ACT1_DROME) and mutant actin (actin NP) were produced and purified from SF9 insect cells infected with recombinant Baculoviruses as described before [ ].

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added. .. To monitor DNA digestion, DNA was extracted from a portion of the extracts using spin column purification (Macherey Nagel) and separated by electrophoresis on a 1% GelRed-agarose gel.

Article Title: Characterization of the Single Stranded DNA Binding Protein SsbB Encoded in the Gonoccocal Genetic Island
Article Snippet: Paragraph title: Expression and purification of SsbB ... After thawing, the solution was supplemented with 1 tablet of Protease Inhibitor Cocktail (Roche) and 1 mgr of DNase I (Roche), and the cells were disrupted 3 times in a high-pressure Cell Disrupter (Constant Cell Disruption Systems) at 2.300 bar.

Article Title: Regulation of DNA nucleases by molecular crowding
Article Snippet: .. Purified DNase I for the circular dichroism (CD) studies was purchased from Roche (Mannheim, Germany). ..

Mouse Assay:

Article Title: Neutrophil Extracellular Traps Are Pathogenic in Ventilator-Induced Lung Injury and Partially Dependent on TLR4
Article Snippet: .. Discussion In this study, we illustrated the following: ventilation with high-tidal volume causes lung injury and inflammation in mice; NETs increase in the lung of the VILI animal model upon the elevation of known NET-associated proteins and extracellular DNA; pretreatment of NET degradation with DNase I alleviates the preceding lung injury, which means that NET formation has an adverse effect on the development of VILI; and TLR4 is involved in NET formation in VILI in mice. ..

Functional Assay:

Article Title: The Composition and Structure of Biofilms Developed by Propionibacterium acnes Isolated from Cardiac Pacemaker Devices
Article Snippet: .. In this study, to determine the functional contributions of each ECM component (DNA, protein, and poly-GlcNAc) to biofilm formation, we assessed the effect of DNase I, proteinase K, and dispersin B on the biofilm formation. ..

Binding Assay:

Article Title: The core-independent promoter-specific interaction of primary sigma factor
Article Snippet: .. For binding at 20°C, 0.6 (for G3b promoter) or 0.25 U (for trnS promoter) of DNase I was added; for binding at 25°C, 0.4 U (for G3b promoter) of DNase I was added; for binding at 37°C, 0.25 (for G3b promoter) or 0.1 U (for trnS promoter) of DNase I was added. ..

Size-exclusion Chromatography:

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: .. Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ]. .. D . melanogaster cytoplasmic wild-type (Dm-actin, UNIPROT accession number: ACT1_DROME) and mutant actin (actin NP) were produced and purified from SF9 insect cells infected with recombinant Baculoviruses as described before [ ].

Produced:

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ]. .. D . melanogaster cytoplasmic wild-type (Dm-actin, UNIPROT accession number: ACT1_DROME) and mutant actin (actin NP) were produced and purified from SF9 insect cells infected with recombinant Baculoviruses as described before [ ].

Activation Assay:

Article Title: Differential regulation of actin-activated nucleotidyl cyclase virulence factors by filamentous and globular actin
Article Snippet: All experiments were reproduced with several distinct enzyme preparations of both PaExoY and VnExoY and showed highly consistent results in terms of relative substrate specificities and activation by F versus G actin. .. Commercial DNase I (10104159001, Roche) was further purified on a size-exclusion chromatography column (Superdex 75 16/60, buffer: 0.1 M KCl, 15 mM Hepes pH 7.5, 5 mM CaCl2 , with protease inhibitors) to remove protease contamination present in the commercial DNase reagent. α-actin (UniProt P68135) was prepared from acetone dried powder derived from Oryctolagus cuniculus (Rabbit) back and leg muscles in our laboratory according to the method of Spudich and Watt [ ] as described previously [ ].

Lysis:

Article Title: Identification of Novel Nuclear Factor of Activated T Cell (NFAT)-associated Proteins in T Cells *
Article Snippet: Lysis protocol was adopted from Klenova et al. ( ) and was used for all CoIP experiments, including the CoIP-MS experiments. .. For DNase I digest, a further 3 volumes of dilution buffer were added, and the lysate was supplemented with 5 m m MgCl2 and 1 m m CaCl2 , and DNase I (2000 units, recombinant, Roche Applied Science) was added.

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  • 99
    Roche dnase i
    Impact of <t>DNase</t> I treatment on B . pseudomallei biofilm formation. Static biofilms of B . pseudomallei strains L1, P1 and H777 in LB were treated with DNase I (0.01, 0.1 and 1 U/mL) at 0 h, 24 h or 45 h after inoculation and maintained for up to 48 h. Biofilm formation and eDNA concentration of the 2-day biofilms were assessed using crystal-violet absorbance (OD 620 ) and the QuantiFluor dsDNA System, respectively. DNase I buffer acted as control. Biofilm formation of each strain was examined in eight replicates and eDNA was quantified in duplicates, on three independent occasions. Data represents mean ± SD. * p
    Dnase I, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 2243 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnase i/product/Roche
    Average 99 stars, based on 2243 article reviews
    Price from $9.99 to $1999.99
    dnase i - by Bioz Stars, 2020-04
    99/100 stars
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    Impact of DNase I treatment on B . pseudomallei biofilm formation. Static biofilms of B . pseudomallei strains L1, P1 and H777 in LB were treated with DNase I (0.01, 0.1 and 1 U/mL) at 0 h, 24 h or 45 h after inoculation and maintained for up to 48 h. Biofilm formation and eDNA concentration of the 2-day biofilms were assessed using crystal-violet absorbance (OD 620 ) and the QuantiFluor dsDNA System, respectively. DNase I buffer acted as control. Biofilm formation of each strain was examined in eight replicates and eDNA was quantified in duplicates, on three independent occasions. Data represents mean ± SD. * p

    Journal: PLoS ONE

    Article Title: Extracellular DNA facilitates bacterial adhesion during Burkholderia pseudomallei biofilm formation

    doi: 10.1371/journal.pone.0213288

    Figure Lengend Snippet: Impact of DNase I treatment on B . pseudomallei biofilm formation. Static biofilms of B . pseudomallei strains L1, P1 and H777 in LB were treated with DNase I (0.01, 0.1 and 1 U/mL) at 0 h, 24 h or 45 h after inoculation and maintained for up to 48 h. Biofilm formation and eDNA concentration of the 2-day biofilms were assessed using crystal-violet absorbance (OD 620 ) and the QuantiFluor dsDNA System, respectively. DNase I buffer acted as control. Biofilm formation of each strain was examined in eight replicates and eDNA was quantified in duplicates, on three independent occasions. Data represents mean ± SD. * p

    Article Snippet: DNase I noticeably lowered eDNA concentrations in biofilm if the enzyme was added into the starting inoculum (0 h).

    Techniques: Concentration Assay

    Exogenous chromosomal DNA did not alter either untreated biofilm or DNase I-treated biofilm of B . pseudomallei H777. (A) Amount of 2-day B . pseudomallei H777 biofilm formed in LB, treated with DNase I, supplemented with either salmon sperm DNA (SS DNA) or B . pseudomallei genomic DNA (Bp DNA) compared to the controls. Data represents mean ± SD from three independent experiments. (B) Amount of 2-day B . pseudomallei H777 biofilm formed in LB after treatment with 0.01 U/mL DNase I for 3 h, followed by washing steps to remove DNase, and then supplemented with exogenous salmon sperm DNA or B . pseudomallei genomic DNA. Data represents mean ± SD from three independent experiments. ** p

    Journal: PLoS ONE

    Article Title: Extracellular DNA facilitates bacterial adhesion during Burkholderia pseudomallei biofilm formation

    doi: 10.1371/journal.pone.0213288

    Figure Lengend Snippet: Exogenous chromosomal DNA did not alter either untreated biofilm or DNase I-treated biofilm of B . pseudomallei H777. (A) Amount of 2-day B . pseudomallei H777 biofilm formed in LB, treated with DNase I, supplemented with either salmon sperm DNA (SS DNA) or B . pseudomallei genomic DNA (Bp DNA) compared to the controls. Data represents mean ± SD from three independent experiments. (B) Amount of 2-day B . pseudomallei H777 biofilm formed in LB after treatment with 0.01 U/mL DNase I for 3 h, followed by washing steps to remove DNase, and then supplemented with exogenous salmon sperm DNA or B . pseudomallei genomic DNA. Data represents mean ± SD from three independent experiments. ** p

    Article Snippet: DNase I noticeably lowered eDNA concentrations in biofilm if the enzyme was added into the starting inoculum (0 h).

    Techniques:

    DNase I treatment affects initial attachment and biofilm formation of B . pseudomallei . B . pseudomallei L1, P1 and H777 biofilms were grown in LB at 37°C. The biofilms were treated with DNase I (0.01 U/mL) at 0 h and 24 h post-seeding and maintained until 48 h. (A) CLSM images of DNase I treated biofilm structure and eDNA on coverslips. The 2-day biofilm architecture and quantity of eDNA were examined after staining with FITC-ConA (green) and TOTO-3 (red), respectively. The scale bars indicate 10 μm. The images were taken using a Zeiss 800 CLSM microscope (63× magnification). (B) COMSTAT image analysis of DNase I-treated B . pseudomallei biofilms and eDNA. Data represents mean ± SD of 18 images from three independent experiments. * p

    Journal: PLoS ONE

    Article Title: Extracellular DNA facilitates bacterial adhesion during Burkholderia pseudomallei biofilm formation

    doi: 10.1371/journal.pone.0213288

    Figure Lengend Snippet: DNase I treatment affects initial attachment and biofilm formation of B . pseudomallei . B . pseudomallei L1, P1 and H777 biofilms were grown in LB at 37°C. The biofilms were treated with DNase I (0.01 U/mL) at 0 h and 24 h post-seeding and maintained until 48 h. (A) CLSM images of DNase I treated biofilm structure and eDNA on coverslips. The 2-day biofilm architecture and quantity of eDNA were examined after staining with FITC-ConA (green) and TOTO-3 (red), respectively. The scale bars indicate 10 μm. The images were taken using a Zeiss 800 CLSM microscope (63× magnification). (B) COMSTAT image analysis of DNase I-treated B . pseudomallei biofilms and eDNA. Data represents mean ± SD of 18 images from three independent experiments. * p

    Article Snippet: DNase I noticeably lowered eDNA concentrations in biofilm if the enzyme was added into the starting inoculum (0 h).

    Techniques: Confocal Laser Scanning Microscopy, Staining, Microscopy

    TSA mimics estrogen induced activation of BRCA1 by increasing p300 dependent histone acetylation at the BRCA1 promoter. ( a ) Time course of TFF1 , NBR2 , and BRCA1 expression in MCF-7 cells treated 0–24 h with either E2, E2 + cycloheximide (10 µg ml −1 ), TSA (500 ng ml −1 ), or TSA + cycloheximide as indicated. Error bars represent the s.e.m. for N=2 independent biological replicates. ( b ) DNase I hypersensitivity profile of the BRCA1 promoter and an ( HBB ) locus control from MCF-7 cells treated with either estrogen or TSA. The error bars represent the s.e.m. for N=3 biological replicates. ( c ) Acetylated histone H3, acetylated histone H4, HDAC1, BRCA1, p130, and CtBP ChIP profiles at the BRCA1 promoter in control or MCF-7 cells treated 1 h with 500 ng ml −1 TSA. Error bars represent the s.e.m. for N=2 biological replicates. ( d ) Upper panel: TSA stimulated expression of BRCA1 nascent and mature RNA levels in either control or p300 depleted MCF-7. Error bars represent the s.e.m. for N=2 biological replicates. Lower panel: ChIP enrichment for H3 and H4 histone acetylation at the BRCA1 locus in control versus p300 depleted MCF-7 cells with or without TSA stimulation. Means from N=2 independent biological replicates are shown.

    Journal: Nature structural & molecular biology

    Article Title: Transcriptional regulation of BRCA1 expression by a metabolic switch

    doi: 10.1038/nsmb.1941

    Figure Lengend Snippet: TSA mimics estrogen induced activation of BRCA1 by increasing p300 dependent histone acetylation at the BRCA1 promoter. ( a ) Time course of TFF1 , NBR2 , and BRCA1 expression in MCF-7 cells treated 0–24 h with either E2, E2 + cycloheximide (10 µg ml −1 ), TSA (500 ng ml −1 ), or TSA + cycloheximide as indicated. Error bars represent the s.e.m. for N=2 independent biological replicates. ( b ) DNase I hypersensitivity profile of the BRCA1 promoter and an ( HBB ) locus control from MCF-7 cells treated with either estrogen or TSA. The error bars represent the s.e.m. for N=3 biological replicates. ( c ) Acetylated histone H3, acetylated histone H4, HDAC1, BRCA1, p130, and CtBP ChIP profiles at the BRCA1 promoter in control or MCF-7 cells treated 1 h with 500 ng ml −1 TSA. Error bars represent the s.e.m. for N=2 biological replicates. ( d ) Upper panel: TSA stimulated expression of BRCA1 nascent and mature RNA levels in either control or p300 depleted MCF-7. Error bars represent the s.e.m. for N=2 biological replicates. Lower panel: ChIP enrichment for H3 and H4 histone acetylation at the BRCA1 locus in control versus p300 depleted MCF-7 cells with or without TSA stimulation. Means from N=2 independent biological replicates are shown.

    Article Snippet: For each DNase I digestion, approximately 1×106 nuclei were harvested and resuspended in 200 ul of pre-warmed (37 ° C) Buffer A, supplemented with 6 mM CaCl2 , 75 mM NaCl, and the DNase I (0, 170, 340, and 680 units).

    Techniques: Activation Assay, Expressing, Chromatin Immunoprecipitation

    DNase I treatment of DNA and non-HAd Ft completely ablates the pro-inflammatory cytokine response. (A) Wild-type and AIM2 −/− BMDMs (2.5×10 5 cells/well) were incubated either in the absence or presence of DOTAP and 8 µg/ml of genomic DNA purified from Ft LVS. DNA was either untreated or incubated with DNase I as described in Methods . The levels of the cytokines released after 24 h were determined by CBA or ELISA. *** P

    Journal: PLoS ONE

    Article Title: Discordant Results Obtained with Francisella tularensis during In Vitro and In Vivo Immunological Studies Are Attributable to Compromised Bacterial Structural Integrity

    doi: 10.1371/journal.pone.0058513

    Figure Lengend Snippet: DNase I treatment of DNA and non-HAd Ft completely ablates the pro-inflammatory cytokine response. (A) Wild-type and AIM2 −/− BMDMs (2.5×10 5 cells/well) were incubated either in the absence or presence of DOTAP and 8 µg/ml of genomic DNA purified from Ft LVS. DNA was either untreated or incubated with DNase I as described in Methods . The levels of the cytokines released after 24 h were determined by CBA or ELISA. *** P

    Article Snippet: ***P < 0.001. (All results shown were subjected to One-way ANOVA with Bonferroni's Post-test). (E) & (F) Wild-type and AIM2−/− BMDMs were infected with Ft LVS cultivated in MHB either untreated or treated with DNase I at a MOI of 100.

    Techniques: Incubation, Purification, Crocin Bleaching Assay, Enzyme-linked Immunosorbent Assay

    Applicability of the ECM extraction method to  S . epidermidis ,  E . coli  and  P . aeruginosa  biofilms.A. Extracellular matrices extracted from  S . epidermidis  SE04 by the addition of 1.5 M NaCl were treated with or without dispersin B and were then applied to SDS-APGE. The gel was stained with CBB. An arrow indicates polysaccharides.B. Extracellular matrices of  E . coli  YMel and its isogenic  csgA  mutant YMel-1 were isolated with 1.5 M NaCl. Curli amyloid fibres in the ECM fraction were treated with or without formic acid. Purified curli was also used as a positive control. The proteins were analysed by Western blotting using anti-CsgA antibody. Arrows indicate monomeric and dimeric CsgA. FA, formic acid.C. Extracellular matrices of  P . aeruginosa  PAO1 were extracted with 1.5 M NaCl and were subjected to agarose gel electrophoresis. The extracted ECMs were treated with or without DNase I. The gel was stained with ethidium bromide. The positions of molecular mass markers in kilodaltons (kDa) (A and B) and kilobase pairs (kb) (C) are shown at the left of each panel respectively.

    Journal: Microbial Biotechnology

    Article Title: A refined technique for extraction of extracellular matrices from bacterial biofilms and its applicability

    doi: 10.1111/1751-7915.12155

    Figure Lengend Snippet: Applicability of the ECM extraction method to S . epidermidis , E . coli and P . aeruginosa biofilms.A. Extracellular matrices extracted from S . epidermidis  SE04 by the addition of 1.5 M NaCl were treated with or without dispersin B and were then applied to SDS-APGE. The gel was stained with CBB. An arrow indicates polysaccharides.B. Extracellular matrices of E . coli  YMel and its isogenic csgA mutant YMel-1 were isolated with 1.5 M NaCl. Curli amyloid fibres in the ECM fraction were treated with or without formic acid. Purified curli was also used as a positive control. The proteins were analysed by Western blotting using anti-CsgA antibody. Arrows indicate monomeric and dimeric CsgA. FA, formic acid.C. Extracellular matrices of P . aeruginosa  PAO1 were extracted with 1.5 M NaCl and were subjected to agarose gel electrophoresis. The extracted ECMs were treated with or without DNase I. The gel was stained with ethidium bromide. The positions of molecular mass markers in kilodaltons (kDa) (A and B) and kilobase pairs (kb) (C) are shown at the left of each panel respectively.

    Article Snippet: ECM-degrading enzymes Dispersin B from Aggregatibacter actinomycetemcomitans (20 μg ml−1 , Kane Biotech Inc., Manitoba, Canada), Proteinase K from Tritirachium album (100 μg ml−1 , Sigma, St Louis, MO, USA) and DNase I (100 U ml−1 , Roche Diagnostics, Mannheim, Germany) were used for dispersal of preformed biofilms or degradation of ECM components.

    Techniques: Staining, Mutagenesis, Isolation, Purification, Positive Control, Western Blot, Agarose Gel Electrophoresis