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FUJIFILM dnaase i
Dnaase I, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 3 article reviews
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dnaase i - by Bioz Stars, 2020-04
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Related Articles

Cell Isolation:

Article Title: Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis
Article Snippet: Paragraph title: Cell isolation ... To isolate lamina propria cells, the remaining colons after removing epithelial layer were chopped into little pieces with scalpel and digested for 60 min at 37°C with gentle shaking in RPMI1640 medium supplemented with 5% FBS, 0.5 mg/ml Collagenase IV (SIGMA) and 50 U/ml DNase I (Wako).

Amplification:

Article Title: Localization of an hTERT repressor region on human chromosome 3p21.3 using chromosome engineering
Article Snippet: RNA isolation and RT-PCR for hTERT expression Total RNA was extracted by using RNeasy mini kit (Qiagen) and treated with DNase I (Wako). .. The first-strand cDNA was synthesized using M-MLV reverse transcriptase (Invitrogen) with the oligo(dT)15 primer. cDNA was amplified using 5'-CGA GAG CAG ACA CCA GCA G-3' and 5'-TTTTACTCCCACAGCACCTC-3' for endogenous hTERT ; 5'-GAC GAC GAT GAC AAG GGA AT-3' and 5'-AGC ACC TCG CGG TAG TGG-3' for exogenous hTERT ; 5'-CCA TCT TCC AGG AGC GAG A-3' and 5'-TGT CAT ACC AGG AAA TGA GC-3' for GAPDH .

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo). .. PCR was initiated with DNA polymerase (G-Taq; Cosmo Genetech, Seoul, Korea) at 95°C for 2 min, followed by 35 cycles of amplification at 95°C for 20 s, 56°C for 40 s, 72°C for 10 s, and 72°C for 5 min for final extension.

Article Title: Cytosolic Genomic DNA functions as a Natural Antisense
Article Snippet: For intron PCR, RNA was treated with DNase I (Wako Pure Chemical Industries) for 1 h at 37 °C followed by acid phenol:chloroform extraction. .. All purified DNA were adjusted to a concentration of 6 ng/μl for the PCR amplification.

Synthesized:

Article Title: Localization of an hTERT repressor region on human chromosome 3p21.3 using chromosome engineering
Article Snippet: RNA isolation and RT-PCR for hTERT expression Total RNA was extracted by using RNeasy mini kit (Qiagen) and treated with DNase I (Wako). .. The first-strand cDNA was synthesized using M-MLV reverse transcriptase (Invitrogen) with the oligo(dT)15 primer. cDNA was amplified using 5'-CGA GAG CAG ACA CCA GCA G-3' and 5'-TTTTACTCCCACAGCACCTC-3' for endogenous hTERT ; 5'-GAC GAC GAT GAC AAG GGA AT-3' and 5'-AGC ACC TCG CGG TAG TGG-3' for exogenous hTERT ; 5'-CCA TCT TCC AGG AGC GAG A-3' and 5'-TGT CAT ACC AGG AAA TGA GC-3' for GAPDH .

Cytometry:

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: The population of GFP-positive blood cells prepared from GM chickens was measured by flow cytometry (BD FACSCalibur™). .. Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo).

Article Title: Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity
Article Snippet: .. Flow cytometry Colons, distal Peyer’s patches (ileal Peyer’s patches), and cecal patches were dissociated with RPMI 1640 medium containing 12.5 mM HEPES (pH 7.4), 2% fetal bovine serum, 0.5 mg/ml collagenase (Wako Pure Chemical Industries), and 0.5 mg/ml DNase I (Wako Pure Chemical Industries) at 37 °C for 30 min . .. Cells were isolated by Percoll gradient separation.

Blocking Assay:

Article Title: Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity
Article Snippet: Flow cytometry Colons, distal Peyer’s patches (ileal Peyer’s patches), and cecal patches were dissociated with RPMI 1640 medium containing 12.5 mM HEPES (pH 7.4), 2% fetal bovine serum, 0.5 mg/ml collagenase (Wako Pure Chemical Industries), and 0.5 mg/ml DNase I (Wako Pure Chemical Industries) at 37 °C for 30 min . .. Isolated cells were incubated with anti-CD16/CD32 (FcγR) antibody (catalog number 101319, BioLegend, dilution 1:500) to block non-specific reactions and subsequently stained by using specific antibodies.

Enzyme-linked Immunosorbent Assay:

Article Title: Periodontitis induced by bacterial infection exacerbates features of Alzheimer’s disease in transgenic mice
Article Snippet: The cortical fragments were incubated with 0.25% trypsin and 20 µg/ml DNase I in phosphate-buffered saline at 37 °C for 15 min, dissociated into single cells by pipetting, and then resuspended in serum-free Dulbecco’s modified Eagle’s medium/Ham’s F-12 (Wako Pure Chemical Industries, Osaka, Japan) supplemented with N2 Supplement with Transferrin (Wako Pure Chemical Industries, Osaka, Japan) supplemented with N2 Supplement with Transferrin (Wako Pure Chemical Industries, Osaka, Japan). .. The neurons were stimulated with or without 0.1, 1, 10 µg/ml of P. gingivalis -derived LPS for 24 h. Then levels of Aβ40 and Aβ42 in the media were measured by an ELISA (Wako Pure Chemical Industries).

Incubation:

Article Title: Demonstration of Hepatitis C Virus RNA with In Situ Hybridization Employing a Locked Nucleic Acid Probe in Humanized Liver of Infected Chimeric Mice and in Needle-Biopsied Human Liver
Article Snippet: .. Namely, either 10 IU/mL DNase I (Wako, Osaka, Japan) or 100 μ g/mL RNase (Sigma-Aldrich, St. Louis, MO, USA) was incubated for 30 minutes at 37°C. .. Immunostaining Using a Human Specific Cytokeratin 8/18 (CK8/18) Antibody In order to distinguish mouse hepatocytes from chimeric human hepatocytes in HCV-infected chimeric mice, we utilized a human-specific CK8/18 monoclonal antibody (clone: NCL 5D3, MP Biochemicals, Santa Ana, CA, USA).

Article Title: Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis
Article Snippet: Isolated colons were cut, opened longitudinally and washed with excess PBS to remove stools and mucus, followed by incubation for 30 min at 37°C with vigorous shaking in Ca2+ - and Mg2+ -free Hanks balanced salt solution supplemented with 5 mM EDTA and 1 mM DTT to remove epithelial layer. .. To isolate lamina propria cells, the remaining colons after removing epithelial layer were chopped into little pieces with scalpel and digested for 60 min at 37°C with gentle shaking in RPMI1640 medium supplemented with 5% FBS, 0.5 mg/ml Collagenase IV (SIGMA) and 50 U/ml DNase I (Wako).

Article Title: Periodontitis induced by bacterial infection exacerbates features of Alzheimer’s disease in transgenic mice
Article Snippet: .. The cortical fragments were incubated with 0.25% trypsin and 20 µg/ml DNase I in phosphate-buffered saline at 37 °C for 15 min, dissociated into single cells by pipetting, and then resuspended in serum-free Dulbecco’s modified Eagle’s medium/Ham’s F-12 (Wako Pure Chemical Industries, Osaka, Japan) supplemented with N2 Supplement with Transferrin (Wako Pure Chemical Industries, Osaka, Japan) supplemented with N2 Supplement with Transferrin (Wako Pure Chemical Industries, Osaka, Japan). ..

Article Title: Epithelial‐mesenchymal transition is activated in CD44‐positive malignant ascites tumor cells of gastrointestinal cancer, et al. Epithelial‐mesenchymal transition is activated in CD44‐positive malignant ascites tumor cells of gastrointestinal cancer
Article Snippet: .. 2.3 Primary tissues Surgical resected tumor specimens were washed twice in PBS, minced with a razor blade, and incubated with type 3 collagenase (Worthington Biochemicals, Lakewood, NJ, USA) and DNAase I (Wako Chemicals) for 60 minutes at 37°C using the gentleMACS Dissociator. .. After enzymatic digestion, the sample was sequentially filtered through a 100 μmol/L and 40 μm cell strainer (Corning, Corning, NY, USA), and the cells were incubated with BD Pharm Lyse Lysing Buffer (BD Biosciences) for 5 minutes to lyse red blood cells.

Article Title: Anti PD-1 treatment increases [18F]FDG uptake by cancer cells in a mouse B16F10 melanoma model
Article Snippet: Flow-cytometry analysis Tumors were harvested and processed using Collagenase I and DNase I (Wako, Osaka, Japan). .. Splenocytes were hemolyzed and incubated with anti-CD16/32 2.4G2 (BD Biosciences, San Jose, CA, USA) to reduce FcγR binding.

Article Title: Neutrophils Play an Essential Role in Cooperation with Antibody in both Protection against and Recovery from Pulmonary Infection with Influenza Virus in Mice ▿
Article Snippet: .. Freshly resected lungs were minced with scissors to a fine slurry and enzymatically digested by incubation at 37°C for 1 h in supplemented RPMI 1640 medium (15 ml/lung) containing type A collagenase (1 mg/ml; Wako Pure Chemicals Co., Osaka, Japan) and DNase I (30 U/ml; Wako Pure Chemicals). ..

Article Title: Cytosolic Genomic DNA functions as a Natural Antisense
Article Snippet: The nuclear, mitochondrial, and cytoplasmic fractions were incubated in lysis buffer for 1 h at 37 °C for the further extraction, as previously reported with minor modifications (50 mM HEPES, pH 8.0, 150 mM NaCl, 10% glycerol, 1 mM DTT, 2 mM EDTA, 0.5% SDS, and 100 μg/mL proteinase K (Sigma-Aldrich)). .. For intron PCR, RNA was treated with DNase I (Wako Pure Chemical Industries) for 1 h at 37 °C followed by acid phenol:chloroform extraction.

Article Title: Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater
Article Snippet: .. One microliter of DNase I (Wako, Jun-yaku, Osaka, Japan) and 1.5 μl of 5× DNase I buffer were added to the extracted RNA samples and incubated for 30 min at 37°C, followed by inactivation of DNase I at 75°C for 5 min, and then cooling down to 4°C. .. An RNA-extracted sample was added to 1.5 μl of SuperScript II (Life Technologies Japan, Tokyo, Japan), 1.5 μl of 100 mM dithiothreitol, 6.0 μl of 5× first-strand buffer, 1.5 μl of deoxynucleoside triphosphates (10 mM each), 0.75 μl of RNase inhibitor (20 U/liter; Life Technologies Japan), 0.75 μl of 0.1 M random hexamer, and distilled water to obtain a final volume of 30 μl; incubated at 42°C for 60 min followed by 99°C for 5 min; and then cooled down to 4°C.

Article Title: Control of protein function through oxidation and reduction of persulfidated states
Article Snippet: After peroxide treatment, cells were washed in PBS three times and lysed in 1 ml of lysis buffer [10 mM tris-HCl (pH 7.4), 150 mM NaCl, 0.1% NP-40, 0.1% sodium deoxycholate, deoxyribonuclease I (10 μg/ml), ribonuclease A (10 μg/ml), and protease inhibitor cocktail] without SDS containing 5 mM iodoacetamide (Fujifilm Wako Pure Chemical Corporation). .. The samples were incubated with 50 μl of lysis buffer–washed protein G Mag beads for 2 hours at 4°C.

Article Title: Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity
Article Snippet: Flow cytometry Colons, distal Peyer’s patches (ileal Peyer’s patches), and cecal patches were dissociated with RPMI 1640 medium containing 12.5 mM HEPES (pH 7.4), 2% fetal bovine serum, 0.5 mg/ml collagenase (Wako Pure Chemical Industries), and 0.5 mg/ml DNase I (Wako Pure Chemical Industries) at 37 °C for 30 min . .. Isolated cells were incubated with anti-CD16/CD32 (FcγR) antibody (catalog number 101319, BioLegend, dilution 1:500) to block non-specific reactions and subsequently stained by using specific antibodies.

Activity Assay:

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: POD activity was detected using an enhanced chemiluminescence kit (ECL Plus Western Blotting Detection System; GE Healthcare). .. Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo).

Cell Culture:

Article Title: Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater
Article Snippet: Paragraph title: Determination of occurrence of viruses by cell culture RT-PCR and by direct RT-PCR. ... One microliter of DNase I (Wako, Jun-yaku, Osaka, Japan) and 1.5 μl of 5× DNase I buffer were added to the extracted RNA samples and incubated for 30 min at 37°C, followed by inactivation of DNase I at 75°C for 5 min, and then cooling down to 4°C.

Article Title: Control of protein function through oxidation and reduction of persulfidated states
Article Snippet: Detection of Prx2 Cp modifications in HEK293 cells Normal control (NC) and TrxR1 KD HEK293 cells were seeded to cell culture dishes at 3 × 106 cells per 100-mm2 density 48 hours before experiments. .. After peroxide treatment, cells were washed in PBS three times and lysed in 1 ml of lysis buffer [10 mM tris-HCl (pH 7.4), 150 mM NaCl, 0.1% NP-40, 0.1% sodium deoxycholate, deoxyribonuclease I (10 μg/ml), ribonuclease A (10 μg/ml), and protease inhibitor cocktail] without SDS containing 5 mM iodoacetamide (Fujifilm Wako Pure Chemical Corporation).

Article Title: Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity
Article Snippet: Flow cytometry Colons, distal Peyer’s patches (ileal Peyer’s patches), and cecal patches were dissociated with RPMI 1640 medium containing 12.5 mM HEPES (pH 7.4), 2% fetal bovine serum, 0.5 mg/ml collagenase (Wako Pure Chemical Industries), and 0.5 mg/ml DNase I (Wako Pure Chemical Industries) at 37 °C for 30 min . .. For intracellular cytokine staining, isolated cells were cultured with phorbol 12-myristate 13-acetate, ionomycin, and brefeldin A for 4.5 h prior to cell surface staining.

Expressing:

Article Title: Localization of an hTERT repressor region on human chromosome 3p21.3 using chromosome engineering
Article Snippet: .. RNA isolation and RT-PCR for hTERT expression Total RNA was extracted by using RNeasy mini kit (Qiagen) and treated with DNase I (Wako). .. The first-strand cDNA was synthesized using M-MLV reverse transcriptase (Invitrogen) with the oligo(dT)15 primer. cDNA was amplified using 5'-CGA GAG CAG ACA CCA GCA G-3' and 5'-TTTTACTCCCACAGCACCTC-3' for endogenous hTERT ; 5'-GAC GAC GAT GAC AAG GGA AT-3' and 5'-AGC ACC TCG CGG TAG TGG-3' for exogenous hTERT ; 5'-CCA TCT TCC AGG AGC GAG A-3' and 5'-TGT CAT ACC AGG AAA TGA GC-3' for GAPDH .

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: Paragraph title: Transgene expression analysis ... Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo).

Modification:

Article Title: Periodontitis induced by bacterial infection exacerbates features of Alzheimer’s disease in transgenic mice
Article Snippet: .. The cortical fragments were incubated with 0.25% trypsin and 20 µg/ml DNase I in phosphate-buffered saline at 37 °C for 15 min, dissociated into single cells by pipetting, and then resuspended in serum-free Dulbecco’s modified Eagle’s medium/Ham’s F-12 (Wako Pure Chemical Industries, Osaka, Japan) supplemented with N2 Supplement with Transferrin (Wako Pure Chemical Industries, Osaka, Japan) supplemented with N2 Supplement with Transferrin (Wako Pure Chemical Industries, Osaka, Japan). ..

Western Blot:

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: POD activity was detected using an enhanced chemiluminescence kit (ECL Plus Western Blotting Detection System; GE Healthcare). .. Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo).

Derivative Assay:

Article Title: Periodontitis induced by bacterial infection exacerbates features of Alzheimer’s disease in transgenic mice
Article Snippet: The cortical fragments were incubated with 0.25% trypsin and 20 µg/ml DNase I in phosphate-buffered saline at 37 °C for 15 min, dissociated into single cells by pipetting, and then resuspended in serum-free Dulbecco’s modified Eagle’s medium/Ham’s F-12 (Wako Pure Chemical Industries, Osaka, Japan) supplemented with N2 Supplement with Transferrin (Wako Pure Chemical Industries, Osaka, Japan) supplemented with N2 Supplement with Transferrin (Wako Pure Chemical Industries, Osaka, Japan). .. The neurons were stimulated with or without 0.1, 1, 10 µg/ml of P. gingivalis -derived LPS for 24 h. Then levels of Aβ40 and Aβ42 in the media were measured by an ELISA (Wako Pure Chemical Industries).

Hybridization:

Article Title: Demonstration of Hepatitis C Virus RNA with In Situ Hybridization Employing a Locked Nucleic Acid Probe in Humanized Liver of Infected Chimeric Mice and in Needle-Biopsied Human Liver
Article Snippet: In order to verify the specificity of the ISH technique for HCV-RNA detection, we carried out the treatment with DNase and RNase before the hybridization with the LNA probe on consecutive sections. .. Namely, either 10 IU/mL DNase I (Wako, Osaka, Japan) or 100 μ g/mL RNase (Sigma-Aldrich, St. Louis, MO, USA) was incubated for 30 minutes at 37°C.

Flow Cytometry:

Article Title: Anti PD-1 treatment increases [18F]FDG uptake by cancer cells in a mouse B16F10 melanoma model
Article Snippet: .. Flow-cytometry analysis Tumors were harvested and processed using Collagenase I and DNase I (Wako, Osaka, Japan). ..

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: The population of GFP-positive blood cells prepared from GM chickens was measured by flow cytometry (BD FACSCalibur™). .. Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo).

Article Title: Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity
Article Snippet: .. Flow cytometry Colons, distal Peyer’s patches (ileal Peyer’s patches), and cecal patches were dissociated with RPMI 1640 medium containing 12.5 mM HEPES (pH 7.4), 2% fetal bovine serum, 0.5 mg/ml collagenase (Wako Pure Chemical Industries), and 0.5 mg/ml DNase I (Wako Pure Chemical Industries) at 37 °C for 30 min . .. Cells were isolated by Percoll gradient separation.

Protease Inhibitor:

Article Title: Control of protein function through oxidation and reduction of persulfidated states
Article Snippet: .. After peroxide treatment, cells were washed in PBS three times and lysed in 1 ml of lysis buffer [10 mM tris-HCl (pH 7.4), 150 mM NaCl, 0.1% NP-40, 0.1% sodium deoxycholate, deoxyribonuclease I (10 μg/ml), ribonuclease A (10 μg/ml), and protease inhibitor cocktail] without SDS containing 5 mM iodoacetamide (Fujifilm Wako Pure Chemical Corporation). .. Supernatants were transferred into low protein–binding microcentrifuge tubes (LoBind Tubes, Eppendorf), mixed with 30 μl of protein G Mag beads (GE Healthcare; previously washed with lysis buffer), and agitated for 20 min at 4°C.

Infection:

Article Title: Neutrophils Play an Essential Role in Cooperation with Antibody in both Protection against and Recovery from Pulmonary Infection with Influenza Virus in Mice ▿
Article Snippet: Paragraph title: Preparation of cells infiltrating the lungs of mice infected with influenza virus. ... Freshly resected lungs were minced with scissors to a fine slurry and enzymatically digested by incubation at 37°C for 1 h in supplemented RPMI 1640 medium (15 ml/lung) containing type A collagenase (1 mg/ml; Wako Pure Chemicals Co., Osaka, Japan) and DNase I (30 U/ml; Wako Pure Chemicals).

Polymerase Chain Reaction:

Article Title: Localization of an hTERT repressor region on human chromosome 3p21.3 using chromosome engineering
Article Snippet: RNA isolation and RT-PCR for hTERT expression Total RNA was extracted by using RNeasy mini kit (Qiagen) and treated with DNase I (Wako). .. The PCR reaction for endogenous hTERT was performed using Advantage2 PCR kit (Clontech).

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo). .. The cDNAs from each sample were subjected to PCR.

Article Title: Cytosolic Genomic DNA functions as a Natural Antisense
Article Snippet: .. For intron PCR, RNA was treated with DNase I (Wako Pure Chemical Industries) for 1 h at 37 °C followed by acid phenol:chloroform extraction. .. For PCR of cytosolic genomic DNA PCR, DNA was treated with an RNase cocktail (Thermo Fisher Scientific) for 1 h at 37 °C followed by PCI extraction.

Article Title: Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater
Article Snippet: One microliter of DNase I (Wako, Jun-yaku, Osaka, Japan) and 1.5 μl of 5× DNase I buffer were added to the extracted RNA samples and incubated for 30 min at 37°C, followed by inactivation of DNase I at 75°C for 5 min, and then cooling down to 4°C. .. PCR was performed using a TaqMan PCR reagent kit (PE Biosystems Japan, Urayasu, Japan) with a 200 nM concentration of each primer and a 100 nM concentration of TaqMan probe.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Localization of an hTERT repressor region on human chromosome 3p21.3 using chromosome engineering
Article Snippet: .. RNA isolation and RT-PCR for hTERT expression Total RNA was extracted by using RNeasy mini kit (Qiagen) and treated with DNase I (Wako). .. The first-strand cDNA was synthesized using M-MLV reverse transcriptase (Invitrogen) with the oligo(dT)15 primer. cDNA was amplified using 5'-CGA GAG CAG ACA CCA GCA G-3' and 5'-TTTTACTCCCACAGCACCTC-3' for endogenous hTERT ; 5'-GAC GAC GAT GAC AAG GGA AT-3' and 5'-AGC ACC TCG CGG TAG TGG-3' for exogenous hTERT ; 5'-CCA TCT TCC AGG AGC GAG A-3' and 5'-TGT CAT ACC AGG AAA TGA GC-3' for GAPDH .

Article Title: Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater
Article Snippet: Paragraph title: Determination of occurrence of viruses by cell culture RT-PCR and by direct RT-PCR. ... One microliter of DNase I (Wako, Jun-yaku, Osaka, Japan) and 1.5 μl of 5× DNase I buffer were added to the extracted RNA samples and incubated for 30 min at 37°C, followed by inactivation of DNase I at 75°C for 5 min, and then cooling down to 4°C.

Injection:

Article Title: Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis
Article Snippet: Cell isolation Mice were euthanized by intraperitoneal injection of a large excess of pentobarbital sodium. .. To isolate lamina propria cells, the remaining colons after removing epithelial layer were chopped into little pieces with scalpel and digested for 60 min at 37°C with gentle shaking in RPMI1640 medium supplemented with 5% FBS, 0.5 mg/ml Collagenase IV (SIGMA) and 50 U/ml DNase I (Wako).

Binding Assay:

Article Title: Anti PD-1 treatment increases [18F]FDG uptake by cancer cells in a mouse B16F10 melanoma model
Article Snippet: Flow-cytometry analysis Tumors were harvested and processed using Collagenase I and DNase I (Wako, Osaka, Japan). .. Splenocytes were hemolyzed and incubated with anti-CD16/32 2.4G2 (BD Biosciences, San Jose, CA, USA) to reduce FcγR binding.

Cellular Antioxidant Activity Assay:

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo). .. The following primers were used; 5′-ACC GTA ACA CCG ATG GGA GTA-3′ and 5′-ACA CCG GCC TTA TTC CAA-3′ for the cLys-7crp sequence and 5′-CAC AGA AGA CGG TGG ATG GC-3′ and 5′-GAG ACA TTG GGG GTT GGC A-3′ for the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene.

Isolation:

Article Title: Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis
Article Snippet: Isolated colons were cut, opened longitudinally and washed with excess PBS to remove stools and mucus, followed by incubation for 30 min at 37°C with vigorous shaking in Ca2+ - and Mg2+ -free Hanks balanced salt solution supplemented with 5 mM EDTA and 1 mM DTT to remove epithelial layer. .. To isolate lamina propria cells, the remaining colons after removing epithelial layer were chopped into little pieces with scalpel and digested for 60 min at 37°C with gentle shaking in RPMI1640 medium supplemented with 5% FBS, 0.5 mg/ml Collagenase IV (SIGMA) and 50 U/ml DNase I (Wako).

Article Title: Localization of an hTERT repressor region on human chromosome 3p21.3 using chromosome engineering
Article Snippet: .. RNA isolation and RT-PCR for hTERT expression Total RNA was extracted by using RNeasy mini kit (Qiagen) and treated with DNase I (Wako). .. The first-strand cDNA was synthesized using M-MLV reverse transcriptase (Invitrogen) with the oligo(dT)15 primer. cDNA was amplified using 5'-CGA GAG CAG ACA CCA GCA G-3' and 5'-TTTTACTCCCACAGCACCTC-3' for endogenous hTERT ; 5'-GAC GAC GAT GAC AAG GGA AT-3' and 5'-AGC ACC TCG CGG TAG TGG-3' for exogenous hTERT ; 5'-CCA TCT TCC AGG AGC GAG A-3' and 5'-TGT CAT ACC AGG AAA TGA GC-3' for GAPDH .

Article Title: Neutrophils Play an Essential Role in Cooperation with Antibody in both Protection against and Recovery from Pulmonary Infection with Influenza Virus in Mice ▿
Article Snippet: Freshly resected lungs were minced with scissors to a fine slurry and enzymatically digested by incubation at 37°C for 1 h in supplemented RPMI 1640 medium (15 ml/lung) containing type A collagenase (1 mg/ml; Wako Pure Chemicals Co., Osaka, Japan) and DNase I (30 U/ml; Wako Pure Chemicals). .. The isolated cells were counted using a hemocytometer, and cell suspensions were first applied to glass slides and then dried and fixed with methanol and stained with Giemsa.

Article Title: Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity
Article Snippet: Flow cytometry Colons, distal Peyer’s patches (ileal Peyer’s patches), and cecal patches were dissociated with RPMI 1640 medium containing 12.5 mM HEPES (pH 7.4), 2% fetal bovine serum, 0.5 mg/ml collagenase (Wako Pure Chemical Industries), and 0.5 mg/ml DNase I (Wako Pure Chemical Industries) at 37 °C for 30 min . .. Cells were isolated by Percoll gradient separation.

Microscopy:

Article Title: Neutrophils Play an Essential Role in Cooperation with Antibody in both Protection against and Recovery from Pulmonary Infection with Influenza Virus in Mice ▿
Article Snippet: Freshly resected lungs were minced with scissors to a fine slurry and enzymatically digested by incubation at 37°C for 1 h in supplemented RPMI 1640 medium (15 ml/lung) containing type A collagenase (1 mg/ml; Wako Pure Chemicals Co., Osaka, Japan) and DNase I (30 U/ml; Wako Pure Chemicals). .. The differential cell counts were determined under a microscope, and the absolute number of a leukocyte subtype was determined by multiplication of the percentage of that cell type by the total number of lung leukocytes in the sample.

Mouse Assay:

Article Title: Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis
Article Snippet: Cell isolation Mice were euthanized by intraperitoneal injection of a large excess of pentobarbital sodium. .. To isolate lamina propria cells, the remaining colons after removing epithelial layer were chopped into little pieces with scalpel and digested for 60 min at 37°C with gentle shaking in RPMI1640 medium supplemented with 5% FBS, 0.5 mg/ml Collagenase IV (SIGMA) and 50 U/ml DNase I (Wako).

Article Title: Neutrophils Play an Essential Role in Cooperation with Antibody in both Protection against and Recovery from Pulmonary Infection with Influenza Virus in Mice ▿
Article Snippet: Paragraph title: Preparation of cells infiltrating the lungs of mice infected with influenza virus. ... Freshly resected lungs were minced with scissors to a fine slurry and enzymatically digested by incubation at 37°C for 1 h in supplemented RPMI 1640 medium (15 ml/lung) containing type A collagenase (1 mg/ml; Wako Pure Chemicals Co., Osaka, Japan) and DNase I (30 U/ml; Wako Pure Chemicals).

Sequencing:

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo). .. The following primers were used; 5′-ACC GTA ACA CCG ATG GGA GTA-3′ and 5′-ACA CCG GCC TTA TTC CAA-3′ for the cLys-7crp sequence and 5′-CAC AGA AGA CGG TGG ATG GC-3′ and 5′-GAG ACA TTG GGG GTT GGC A-3′ for the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene.

Staining:

Article Title: Anti PD-1 treatment increases [18F]FDG uptake by cancer cells in a mouse B16F10 melanoma model
Article Snippet: Flow-cytometry analysis Tumors were harvested and processed using Collagenase I and DNase I (Wako, Osaka, Japan). .. Cell-surface antigens were stained with antibodies specific for CD8 (BioLegend, San Diego, CA, USA, clone 53-6.7), CD4 (BioLegend, clone GK1.5), CD45 (BioLegend, clone 30-F11), IA/IE (BD Bioscience, clone M5/114), CD11c (BD Bioscience, clone HL3), CD11b (BioLegend, clone M1/70), and F4/80 (BioLegend, clone BM8).

Article Title: Neutrophils Play an Essential Role in Cooperation with Antibody in both Protection against and Recovery from Pulmonary Infection with Influenza Virus in Mice ▿
Article Snippet: Freshly resected lungs were minced with scissors to a fine slurry and enzymatically digested by incubation at 37°C for 1 h in supplemented RPMI 1640 medium (15 ml/lung) containing type A collagenase (1 mg/ml; Wako Pure Chemicals Co., Osaka, Japan) and DNase I (30 U/ml; Wako Pure Chemicals). .. The isolated cells were counted using a hemocytometer, and cell suspensions were first applied to glass slides and then dried and fixed with methanol and stained with Giemsa.

Article Title: Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity
Article Snippet: Flow cytometry Colons, distal Peyer’s patches (ileal Peyer’s patches), and cecal patches were dissociated with RPMI 1640 medium containing 12.5 mM HEPES (pH 7.4), 2% fetal bovine serum, 0.5 mg/ml collagenase (Wako Pure Chemical Industries), and 0.5 mg/ml DNase I (Wako Pure Chemical Industries) at 37 °C for 30 min . .. Isolated cells were incubated with anti-CD16/CD32 (FcγR) antibody (catalog number 101319, BioLegend, dilution 1:500) to block non-specific reactions and subsequently stained by using specific antibodies.

IA:

Article Title: Anti PD-1 treatment increases [18F]FDG uptake by cancer cells in a mouse B16F10 melanoma model
Article Snippet: Flow-cytometry analysis Tumors were harvested and processed using Collagenase I and DNase I (Wako, Osaka, Japan). .. Cell-surface antigens were stained with antibodies specific for CD8 (BioLegend, San Diego, CA, USA, clone 53-6.7), CD4 (BioLegend, clone GK1.5), CD45 (BioLegend, clone 30-F11), IA/IE (BD Bioscience, clone M5/114), CD11c (BD Bioscience, clone HL3), CD11b (BioLegend, clone M1/70), and F4/80 (BioLegend, clone BM8).

Chloramphenicol Acetyltransferase Assay:

Article Title: Localization of an hTERT repressor region on human chromosome 3p21.3 using chromosome engineering
Article Snippet: RNA isolation and RT-PCR for hTERT expression Total RNA was extracted by using RNeasy mini kit (Qiagen) and treated with DNase I (Wako). .. The first-strand cDNA was synthesized using M-MLV reverse transcriptase (Invitrogen) with the oligo(dT)15 primer. cDNA was amplified using 5'-CGA GAG CAG ACA CCA GCA G-3' and 5'-TTTTACTCCCACAGCACCTC-3' for endogenous hTERT ; 5'-GAC GAC GAT GAC AAG GGA AT-3' and 5'-AGC ACC TCG CGG TAG TGG-3' for exogenous hTERT ; 5'-CCA TCT TCC AGG AGC GAG A-3' and 5'-TGT CAT ACC AGG AAA TGA GC-3' for GAPDH .

Purification:

Article Title: Cytosolic Genomic DNA functions as a Natural Antisense
Article Snippet: For intron PCR, RNA was treated with DNase I (Wako Pure Chemical Industries) for 1 h at 37 °C followed by acid phenol:chloroform extraction. .. All purified DNA were adjusted to a concentration of 6 ng/μl for the PCR amplification.

In Situ Hybridization:

Article Title: Demonstration of Hepatitis C Virus RNA with In Situ Hybridization Employing a Locked Nucleic Acid Probe in Humanized Liver of Infected Chimeric Mice and in Needle-Biopsied Human Liver
Article Snippet: Paragraph title: 2.3. ISH ... Namely, either 10 IU/mL DNase I (Wako, Osaka, Japan) or 100 μ g/mL RNase (Sigma-Aldrich, St. Louis, MO, USA) was incubated for 30 minutes at 37°C.

RNA Extraction:

Article Title: Cytosolic Genomic DNA functions as a Natural Antisense
Article Snippet: Paragraph title: Fractionation and cytosolic DNA and RNA extraction ... For intron PCR, RNA was treated with DNase I (Wako Pure Chemical Industries) for 1 h at 37 °C followed by acid phenol:chloroform extraction.

Low Protein Binding:

Article Title: Control of protein function through oxidation and reduction of persulfidated states
Article Snippet: After peroxide treatment, cells were washed in PBS three times and lysed in 1 ml of lysis buffer [10 mM tris-HCl (pH 7.4), 150 mM NaCl, 0.1% NP-40, 0.1% sodium deoxycholate, deoxyribonuclease I (10 μg/ml), ribonuclease A (10 μg/ml), and protease inhibitor cocktail] without SDS containing 5 mM iodoacetamide (Fujifilm Wako Pure Chemical Corporation). .. Supernatants were transferred into low protein–binding microcentrifuge tubes (LoBind Tubes, Eppendorf), mixed with 30 μl of protein G Mag beads (GE Healthcare; previously washed with lysis buffer), and agitated for 20 min at 4°C.

In Situ:

Article Title: Oral Immunotherapy for Pollen Allergy Using T-Cell Epitope-Containing Egg White Derived from Genetically Manipulated Chickens
Article Snippet: For in situ GFP visualization, a blue light-emitting diode light (cellular blue LED light, LEDB-3WOF; OptoCode, Tokyo, Japan) was shed on the GM chicken through a GFP filter. .. Total RNAs were treated with DNase I (Wako Pure Chemical Industries) and then converted to cDNA using oligo-dT primer and ReverTra Ace reverse transcriptase (Toyobo).

Random Hexamer Labeling:

Article Title: Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater
Article Snippet: One microliter of DNase I (Wako, Jun-yaku, Osaka, Japan) and 1.5 μl of 5× DNase I buffer were added to the extracted RNA samples and incubated for 30 min at 37°C, followed by inactivation of DNase I at 75°C for 5 min, and then cooling down to 4°C. .. An RNA-extracted sample was added to 1.5 μl of SuperScript II (Life Technologies Japan, Tokyo, Japan), 1.5 μl of 100 mM dithiothreitol, 6.0 μl of 5× first-strand buffer, 1.5 μl of deoxynucleoside triphosphates (10 mM each), 0.75 μl of RNase inhibitor (20 U/liter; Life Technologies Japan), 0.75 μl of 0.1 M random hexamer, and distilled water to obtain a final volume of 30 μl; incubated at 42°C for 60 min followed by 99°C for 5 min; and then cooled down to 4°C.

Concentration Assay:

Article Title: Cytosolic Genomic DNA functions as a Natural Antisense
Article Snippet: For intron PCR, RNA was treated with DNase I (Wako Pure Chemical Industries) for 1 h at 37 °C followed by acid phenol:chloroform extraction. .. All purified DNA were adjusted to a concentration of 6 ng/μl for the PCR amplification.

Article Title: Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater
Article Snippet: One microliter of DNase I (Wako, Jun-yaku, Osaka, Japan) and 1.5 μl of 5× DNase I buffer were added to the extracted RNA samples and incubated for 30 min at 37°C, followed by inactivation of DNase I at 75°C for 5 min, and then cooling down to 4°C. .. PCR was performed using a TaqMan PCR reagent kit (PE Biosystems Japan, Urayasu, Japan) with a 200 nM concentration of each primer and a 100 nM concentration of TaqMan probe.

Fractionation:

Article Title: Cytosolic Genomic DNA functions as a Natural Antisense
Article Snippet: Paragraph title: Fractionation and cytosolic DNA and RNA extraction ... For intron PCR, RNA was treated with DNase I (Wako Pure Chemical Industries) for 1 h at 37 °C followed by acid phenol:chloroform extraction.

Lysis:

Article Title: Cytosolic Genomic DNA functions as a Natural Antisense
Article Snippet: The nuclear, mitochondrial, and cytoplasmic fractions were incubated in lysis buffer for 1 h at 37 °C for the further extraction, as previously reported with minor modifications (50 mM HEPES, pH 8.0, 150 mM NaCl, 10% glycerol, 1 mM DTT, 2 mM EDTA, 0.5% SDS, and 100 μg/mL proteinase K (Sigma-Aldrich)). .. For intron PCR, RNA was treated with DNase I (Wako Pure Chemical Industries) for 1 h at 37 °C followed by acid phenol:chloroform extraction.

Article Title: Control of protein function through oxidation and reduction of persulfidated states
Article Snippet: .. After peroxide treatment, cells were washed in PBS three times and lysed in 1 ml of lysis buffer [10 mM tris-HCl (pH 7.4), 150 mM NaCl, 0.1% NP-40, 0.1% sodium deoxycholate, deoxyribonuclease I (10 μg/ml), ribonuclease A (10 μg/ml), and protease inhibitor cocktail] without SDS containing 5 mM iodoacetamide (Fujifilm Wako Pure Chemical Corporation). .. Supernatants were transferred into low protein–binding microcentrifuge tubes (LoBind Tubes, Eppendorf), mixed with 30 μl of protein G Mag beads (GE Healthcare; previously washed with lysis buffer), and agitated for 20 min at 4°C.

Gradient Centrifugation:

Article Title: Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis
Article Snippet: Cells released from colons were subjected to 40% / 70% Percoll gradient centrifugation and the cells containing IELs in middle layer were harvested and used for experiments. .. To isolate lamina propria cells, the remaining colons after removing epithelial layer were chopped into little pieces with scalpel and digested for 60 min at 37°C with gentle shaking in RPMI1640 medium supplemented with 5% FBS, 0.5 mg/ml Collagenase IV (SIGMA) and 50 U/ml DNase I (Wako).

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    FUJIFILM dnase i activity
    Involvement of the amino-acid residues related to each functional SNPs examined in this study on the <t>DNase</t> I activity. Relative activities of amino-acid-substituted constructs related to functional SNPs to that of the wild-type DNase I are presented.
    Dnase I Activity, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 96/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    FUJIFILM dnase i treatment
    BldC binding to the whiI promoter region. a <t>DNase</t> I footprinting analysis. 5′ end-labeled probes were incubated with increasing amounts of BldC (indicated in µM above the lanes) and subjected to DNase I footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced DNase I cleavage on the forward strand and the vertical bar indicates the region of DNase I protection on the reverse strand. b Summary of DNase I footprinting results presented in a . The bracket indicates the protected region and the vertical black arrows indicate sites of enhanced DNase I cleavage. The numbers indicate the distance to the 5′ end of the whiI transcript. The direct repeats shown to be bound by BldC in a head-tail orientation are indicated by arrows (with arrows pointing in the head-tail direction)
    Dnase I Treatment, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    FUJIFILM deoxyribonuclease i
    BldC binding to the whiI promoter region. a <t>DNase</t> I footprinting analysis. 5′ end-labeled probes were incubated with increasing amounts of BldC (indicated in µM above the lanes) and subjected to DNase I footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced DNase I cleavage on the forward strand and the vertical bar indicates the region of DNase I protection on the reverse strand. b Summary of DNase I footprinting results presented in a . The bracket indicates the protected region and the vertical black arrows indicate sites of enhanced DNase I cleavage. The numbers indicate the distance to the 5′ end of the whiI transcript. The direct repeats shown to be bound by BldC in a head-tail orientation are indicated by arrows (with arrows pointing in the head-tail direction)
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    Image Search Results


    Involvement of the amino-acid residues related to each functional SNPs examined in this study on the DNase I activity. Relative activities of amino-acid-substituted constructs related to functional SNPs to that of the wild-type DNase I are presented.

    Journal: DNA and Cell Biology

    Article Title: Identification of the Functional Alleles of the Nonsynonymous Single-Nucleotide Polymorphisms Potentially Implicated in Systemic Lupus Erythematosus in the Human Deoxyribonuclease I Gene

    doi: 10.1089/dna.2014.2368

    Figure Lengend Snippet: Involvement of the amino-acid residues related to each functional SNPs examined in this study on the DNase I activity. Relative activities of amino-acid-substituted constructs related to functional SNPs to that of the wild-type DNase I are presented.

    Article Snippet: The DNase I activity in both the lysates derived from the transfected cells and their conditioned medium was assayed by the single radial enzyme diffusion (SRED) method using a LAS-3000 imaging analyzer (Fuji Film, Tokyo, Japan) according to our previous report (Nadano et al. , ; Yasuda et al. , ).

    Techniques: Functional Assay, Activity Assay, Construct

    Effect of the amino-acid substitution derived from each nonsynonymous SNPs examined in this study on the DNase I activity. The DNase I activities in both (A) the lysates from cells transfected with each construct and (B) their conditioned medium were

    Journal: DNA and Cell Biology

    Article Title: Identification of the Functional Alleles of the Nonsynonymous Single-Nucleotide Polymorphisms Potentially Implicated in Systemic Lupus Erythematosus in the Human Deoxyribonuclease I Gene

    doi: 10.1089/dna.2014.2368

    Figure Lengend Snippet: Effect of the amino-acid substitution derived from each nonsynonymous SNPs examined in this study on the DNase I activity. The DNase I activities in both (A) the lysates from cells transfected with each construct and (B) their conditioned medium were

    Article Snippet: The DNase I activity in both the lysates derived from the transfected cells and their conditioned medium was assayed by the single radial enzyme diffusion (SRED) method using a LAS-3000 imaging analyzer (Fuji Film, Tokyo, Japan) according to our previous report (Nadano et al. , ; Yasuda et al. , ).

    Techniques: Derivative Assay, Activity Assay, Transfection, Construct

    BldC binding to the whiI promoter region. a DNase I footprinting analysis. 5′ end-labeled probes were incubated with increasing amounts of BldC (indicated in µM above the lanes) and subjected to DNase I footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced DNase I cleavage on the forward strand and the vertical bar indicates the region of DNase I protection on the reverse strand. b Summary of DNase I footprinting results presented in a . The bracket indicates the protected region and the vertical black arrows indicate sites of enhanced DNase I cleavage. The numbers indicate the distance to the 5′ end of the whiI transcript. The direct repeats shown to be bound by BldC in a head-tail orientation are indicated by arrows (with arrows pointing in the head-tail direction)

    Journal: Nature Communications

    Article Title: The MerR-like protein BldC binds DNA direct repeats as cooperative multimers to regulate Streptomyces development

    doi: 10.1038/s41467-018-03576-3

    Figure Lengend Snippet: BldC binding to the whiI promoter region. a DNase I footprinting analysis. 5′ end-labeled probes were incubated with increasing amounts of BldC (indicated in µM above the lanes) and subjected to DNase I footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced DNase I cleavage on the forward strand and the vertical bar indicates the region of DNase I protection on the reverse strand. b Summary of DNase I footprinting results presented in a . The bracket indicates the protected region and the vertical black arrows indicate sites of enhanced DNase I cleavage. The numbers indicate the distance to the 5′ end of the whiI transcript. The direct repeats shown to be bound by BldC in a head-tail orientation are indicated by arrows (with arrows pointing in the head-tail direction)

    Article Snippet: Following DNase I treatment and purification, products were separated on denaturing 6% (w/v) polyacrylamide gels and visualized using a FLA-7000 phosphorimager (Fujifilm).

    Techniques: Binding Assay, Footprinting, Labeling, Incubation, Sequencing

    BldC binding to the smeA-ssfA promoter region. a DNase I and b hydroxyl radical footprinting analysis. 5′ end-labeled probes were incubated in the presence (1 µM) or absence of BldC and subjected to footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced cleavage and vertical bars indicate regions of DNase I protection. c Summary of the footprinting results presented in a and b . The filled black circles indicate the repeat pattern of bases protected from hydroxyl radical attack. The bases colored red and blue indicate the DNase I-protected sequences on the forward and reverse strands, respectively, and the vertical black arrows indicate the repeat pattern of DNase I hypersensitive sites seen on the top strand in the presence of BldC. The numbers indicate the distance to the 5′ end of the smeA transcript. Direct repeat motifs are indicated by head-to-tail arrows

    Journal: Nature Communications

    Article Title: The MerR-like protein BldC binds DNA direct repeats as cooperative multimers to regulate Streptomyces development

    doi: 10.1038/s41467-018-03576-3

    Figure Lengend Snippet: BldC binding to the smeA-ssfA promoter region. a DNase I and b hydroxyl radical footprinting analysis. 5′ end-labeled probes were incubated in the presence (1 µM) or absence of BldC and subjected to footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced cleavage and vertical bars indicate regions of DNase I protection. c Summary of the footprinting results presented in a and b . The filled black circles indicate the repeat pattern of bases protected from hydroxyl radical attack. The bases colored red and blue indicate the DNase I-protected sequences on the forward and reverse strands, respectively, and the vertical black arrows indicate the repeat pattern of DNase I hypersensitive sites seen on the top strand in the presence of BldC. The numbers indicate the distance to the 5′ end of the smeA transcript. Direct repeat motifs are indicated by head-to-tail arrows

    Article Snippet: Following DNase I treatment and purification, products were separated on denaturing 6% (w/v) polyacrylamide gels and visualized using a FLA-7000 phosphorimager (Fujifilm).

    Techniques: Binding Assay, Footprinting, Labeling, Incubation, Sequencing

    BldC binding to the whiI promoter region. a DNase I footprinting analysis. 5′ end-labeled probes were incubated with increasing amounts of BldC (indicated in µM above the lanes) and subjected to DNase I footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced DNase I cleavage on the forward strand and the vertical bar indicates the region of DNase I protection on the reverse strand. b Summary of DNase I footprinting results presented in a . The bracket indicates the protected region and the vertical black arrows indicate sites of enhanced DNase I cleavage. The numbers indicate the distance to the 5′ end of the whiI transcript. The direct repeats shown to be bound by BldC in a head-tail orientation are indicated by arrows (with arrows pointing in the head-tail direction)

    Journal: Nature Communications

    Article Title: The MerR-like protein BldC binds DNA direct repeats as cooperative multimers to regulate Streptomyces development

    doi: 10.1038/s41467-018-03576-3

    Figure Lengend Snippet: BldC binding to the whiI promoter region. a DNase I footprinting analysis. 5′ end-labeled probes were incubated with increasing amounts of BldC (indicated in µM above the lanes) and subjected to DNase I footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced DNase I cleavage on the forward strand and the vertical bar indicates the region of DNase I protection on the reverse strand. b Summary of DNase I footprinting results presented in a . The bracket indicates the protected region and the vertical black arrows indicate sites of enhanced DNase I cleavage. The numbers indicate the distance to the 5′ end of the whiI transcript. The direct repeats shown to be bound by BldC in a head-tail orientation are indicated by arrows (with arrows pointing in the head-tail direction)

    Article Snippet: Following DNase I treatment and purification, products were separated on denaturing 6% (w/v) polyacrylamide gels and visualized using a FLA-7000 phosphorimager (Fujifilm).

    Techniques: Binding Assay, Footprinting, Labeling, Incubation, Sequencing

    BldC binding to the smeA-ssfA promoter region. a DNase I and b hydroxyl radical footprinting analysis. 5′ end-labeled probes were incubated in the presence (1 µM) or absence of BldC and subjected to footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced cleavage and vertical bars indicate regions of DNase I protection. c Summary of the footprinting results presented in a and b . The filled black circles indicate the repeat pattern of bases protected from hydroxyl radical attack. The bases colored red and blue indicate the DNase I-protected sequences on the forward and reverse strands, respectively, and the vertical black arrows indicate the repeat pattern of DNase I hypersensitive sites seen on the top strand in the presence of BldC. The numbers indicate the distance to the 5′ end of the smeA transcript. Direct repeat motifs are indicated by head-to-tail arrows

    Journal: Nature Communications

    Article Title: The MerR-like protein BldC binds DNA direct repeats as cooperative multimers to regulate Streptomyces development

    doi: 10.1038/s41467-018-03576-3

    Figure Lengend Snippet: BldC binding to the smeA-ssfA promoter region. a DNase I and b hydroxyl radical footprinting analysis. 5′ end-labeled probes were incubated in the presence (1 µM) or absence of BldC and subjected to footprinting analysis as described in Methods. Footprints are flanked on the left-hand side by Maxam and Gilbert sequence ladders (AG). Horizontal black arrows indicate sites of enhanced cleavage and vertical bars indicate regions of DNase I protection. c Summary of the footprinting results presented in a and b . The filled black circles indicate the repeat pattern of bases protected from hydroxyl radical attack. The bases colored red and blue indicate the DNase I-protected sequences on the forward and reverse strands, respectively, and the vertical black arrows indicate the repeat pattern of DNase I hypersensitive sites seen on the top strand in the presence of BldC. The numbers indicate the distance to the 5′ end of the smeA transcript. Direct repeat motifs are indicated by head-to-tail arrows

    Article Snippet: Following DNase I treatment and purification, products were separated on denaturing 6% (w/v) polyacrylamide gels and visualized using a FLA-7000 phosphorimager (Fujifilm).

    Techniques: Binding Assay, Footprinting, Labeling, Incubation, Sequencing