dnase (New England Biolabs)

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DNase I RNase free

Description:

DNase I RNase free 5 000 units

Catalog Number:

m0303l

Price:

282

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5 000 units

Category:

Deoxyribonucleases DNase

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New England Biolabs dnase

DNase I RNase free 5 000 units
https://www.bioz.com/result/dnase/product/New England Biolabs
Average 99 stars, based on 60 article reviews
Price from $9.99 to $1999.99

dnase - by Bioz Stars, 2020-04

99/100 stars

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Images

1) Product Images from "Holliday junction affinity of the base excision repair factor Endo III contributes to cholera toxin phage integration"

Article Title: Holliday junction affinity of the base excision repair factor Endo III contributes to cholera toxin phage integration

Journal: The EMBO Journal

doi: 10.1038/emboj.2012.219

Endo III blocks XerC-catalysis on pseudo-HJ and displaces it from them. ( A ) Scheme of the suicide pseudo-HJ indicating the mismatch engineered to slow down re-ligation after XerC-cleavage. KMnO 4 sensitive residues in the XerC- and XerD-binding att P(+) arms are indicated by a star. ( B ) Resolution of att P(+)/ dif suicide pseudo-HJs. Legend as in Figure 4D . ( C ) DNase I protection and ( D ) KMnO 4 sensitivity assays of the att P(+)/ dif 1 pseudo-HJ substrate. The analysed strand was labelled on its 5′ end. A scheme of the analysed strand is drawn on the left of the gels. KMnO 4 sensitive residues in the XerC- and XerD-binding sites are indicated by a star.

Figure Legend Snippet: Endo III blocks XerC-catalysis on pseudo-HJ and displaces it from them. ( A ) Scheme of the suicide pseudo-HJ indicating the mismatch engineered to slow down re-ligation after XerC-cleavage. KMnO 4 sensitive residues in the XerC- and XerD-binding att P(+) arms are indicated by a star. ( B ) Resolution of att P(+)/ dif suicide pseudo-HJs. Legend as in Figure 4D . ( C ) DNase I protection and ( D ) KMnO 4 sensitivity assays of the att P(+)/ dif 1 pseudo-HJ substrate. The analysed strand was labelled on its 5′ end. A scheme of the analysed strand is drawn on the left of the gels. KMnO 4 sensitive residues in the XerC- and XerD-binding sites are indicated by a star.

Techniques Used: Ligation, Binding Assay

2) Product Images from "Comparison of Blood RNA Extraction Methods Used for Gene Expression Profiling in Amyotrophic Lateral Sclerosis"

Article Title: Comparison of Blood RNA Extraction Methods Used for Gene Expression Profiling in Amyotrophic Lateral Sclerosis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0087508

Representative Bioanalyzer electropherograms after initial RNA extractions. High quality RNA (RIN > 7.0) can be extracted from all three methods. PAXGENE however requires a DNase step as traces from initial extractions show genomic DNA contamination at high molecular weights and as a “shoulder” to the 28S peak (indicated by asterisks). Good quality RNA can be detected after globin depletion with both TEM and PAXGENE. However, PAXGENE samples showed consistently lower concentration levels, as indicated by the smaller 18S and 28S peaks (FU, fluorescent units). Traces are representative and from single samples from each extraction method.

Figure Legend Snippet: Representative Bioanalyzer electropherograms after initial RNA extractions. High quality RNA (RIN > 7.0) can be extracted from all three methods. PAXGENE however requires a DNase step as traces from initial extractions show genomic DNA contamination at high molecular weights and as a “shoulder” to the 28S peak (indicated by asterisks). Good quality RNA can be detected after globin depletion with both TEM and PAXGENE. However, PAXGENE samples showed consistently lower concentration levels, as indicated by the smaller 18S and 28S peaks (FU, fluorescent units). Traces are representative and from single samples from each extraction method.

Techniques Used: Transmission Electron Microscopy, Concentration Assay

3) Product Images from "3′ IsomiR Species and DNA Contamination Influence Reliable Quantification of MicroRNAs by Stem-Loop Quantitative PCR"

Article Title: 3′ IsomiR Species and DNA Contamination Influence Reliable Quantification of MicroRNAs by Stem-Loop Quantitative PCR

Journal: PLoS ONE

doi: 10.1371/journal.pone.0106315

DNA can serve as a template during miRNA detection. ( A ) False positive signal of DNA derives mainly from the reverse transcription reaction. Mature miR-1226-3p was tested in the indicated samples, with or without reverse transcription (RT). On the y-axis, dC t value is represented, calculated as the Ct difference between the examined samples and the gDNA of control HeLa_mir-33b cell line (Ct = 35,9). One C t difference represents about 2× higher detected mature miRNA level. ( B ) DNA contamination remains in total RNA samples during isolation by the widely used Trizol reagent. Total RNA samples were isolated from transiently transfected HeLa cells; the transfected plasmid DNA amounts are indicated. Samples were DNase treated and non-treated, then reverse transcribed and subjected to real-time PCR. Expression values relative to U6 snRNA are shown on the y-axis. Experiments were carried out with three replicates at least from three independent experiments; one representative experiment is shown, error bars represent standard deviations.

Figure Legend Snippet: DNA can serve as a template during miRNA detection. ( A ) False positive signal of DNA derives mainly from the reverse transcription reaction. Mature miR-1226-3p was tested in the indicated samples, with or without reverse transcription (RT). On the y-axis, dC t value is represented, calculated as the Ct difference between the examined samples and the gDNA of control HeLa_mir-33b cell line (Ct = 35,9). One C t difference represents about 2× higher detected mature miRNA level. ( B ) DNA contamination remains in total RNA samples during isolation by the widely used Trizol reagent. Total RNA samples were isolated from transiently transfected HeLa cells; the transfected plasmid DNA amounts are indicated. Samples were DNase treated and non-treated, then reverse transcribed and subjected to real-time PCR. Expression values relative to U6 snRNA are shown on the y-axis. Experiments were carried out with three replicates at least from three independent experiments; one representative experiment is shown, error bars represent standard deviations.

Techniques Used: Isolation, Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Expressing

Centrifugation:

Article Title: Human U6 promoter drives stronger shRNA activity than its schistosome orthologue in Schistosoma mansoni and human fibrosarcoma cells
Article Snippet: .. Culture supernatants were collected 48 hours later, filtered through 0.45 μm pore size membranes, the filtrate incubated with DNAse I (New England Biolabs, Ipswich, MA) to remove contaminating plasmids, and virions in culture supernatants recovered by centrifugation (Sorvall SS34 rotor, 50,000 g, 90 minutes, 4°C). .. Pelleted virions were resuspended in schistosomule wash medium (above), virion titers estimated using quantitative PCR , and aliquots of virions stored at -80°C until needed. pLNHXΔD70 was employed to produce control virions without shRNA encoding cassettes.

Amplification:

Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
Article Snippet: 188-bp pucB promoter region and 194-bp crtI promoter region containing two TGT-N12 -ACA PpsR-binding palindromes were amplified using a 6-FAM-labeled primer pucBfor ( 5’-AGAGCTCCCGAGGCCCGC-3’ ) and a 5-HEX-labeled primer pucBrev ( 5’-ACCAGTTTCCGTGCTCGACC-3’ ), a 6-FAM-labeled primer crtIfor ( 5’-ATCGGAGGGAACCAGGTGAT-3’ ) and a 5-HEX-labeled primer crtIrev ( 5’-ATCCTGCTTTCCGGCCGCG-3’ ), respectively. .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion.

Article Title: Chromatin remodeling mediated by the FOXA1/A2 transcription factors activates CFTR expression in intestinal epithelial cells
Article Snippet: .. DNase I sensitivity was quantified relative to undigested DNA for each primer set, and shown relative to digestion of a non-DNase I hypersensitive amplicon, DHS1+5 kb, for each DNase I sample. .. The intron 3 (405 + 13.1 kb), DHS10(a,b), and DHS11 fragments were PCR amplified and cloned into the pGL3B luciferase vector (Promega) containing the 787 bp minimal CFTR promoter , using primers listed in , and sequence verified.

Synthesized:

Article Title: Generation of a Novel Nucleic Acid-Based Reporter System To Detect Phenotypic Susceptibility to Antibiotics in Mycobacterium tuberculosis
Article Snippet: BssHII, NotI, XbaI, Antarctic phosphatase, MRI, avian myeloblastosis virus reverse transcriptase, DNase I, and T4 DNA ligase were obtained from New England Biolabs. .. Custom oligodeoxyribonucleotides were synthesized by Eurofins MWG/Operon.

Lambda DNA Preparation:

Article Title: Multipronged regulatory functions of a novel endonuclease (TieA) from Helicobacter pylori
Article Snippet: .. TieA showed a non-specific endonuclease activity similar to DNase I as it cleaved both circular pUC19 and linearized lambda DNA, whereas Rv3131 (lane 6) as well as heat-inactivated TieA (lane 4) did not show any detectable DNA cleavage activity (Figure ). .. Furthermore, our results demonstrated a Ca2+ –Mg2+ dependent DNA cleavage activity of TieA, since EDTA and SDS markedly inhibited the cleavage activity of TieA (Figure , lanes 8, 9, 17, 18).

Construct:

Article Title: Human U6 promoter drives stronger shRNA activity than its schistosome orthologue in Schistosoma mansoni and human fibrosarcoma cells
Article Snippet: Plasmid constructs, prepared with a maxiprep kit (5 Prime, Gaithersburg, MD) were employed to produce the virions as described ( ; ). .. Culture supernatants were collected 48 hours later, filtered through 0.45 μm pore size membranes, the filtrate incubated with DNAse I (New England Biolabs, Ipswich, MA) to remove contaminating plasmids, and virions in culture supernatants recovered by centrifugation (Sorvall SS34 rotor, 50,000 g, 90 minutes, 4°C).

Real-time Polymerase Chain Reaction:

Article Title: Human U6 promoter drives stronger shRNA activity than its schistosome orthologue in Schistosoma mansoni and human fibrosarcoma cells
Article Snippet: Culture supernatants were collected 48 hours later, filtered through 0.45 μm pore size membranes, the filtrate incubated with DNAse I (New England Biolabs, Ipswich, MA) to remove contaminating plasmids, and virions in culture supernatants recovered by centrifugation (Sorvall SS34 rotor, 50,000 g, 90 minutes, 4°C). .. Pelleted virions were resuspended in schistosomule wash medium (above), virion titers estimated using quantitative PCR , and aliquots of virions stored at -80°C until needed. pLNHXΔD70 was employed to produce control virions without shRNA encoding cassettes.

Incubation:

Article Title: A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA
Article Snippet: Pellets were suspended in 50 µl NEB buffer 4 and 10 U Taq I (NEB) and were allowed to incubate at 65°C for 16 h. Taq I was inactivated by heating to 80°C for 20 min followed by an incubation with 2.5 U of shrimp alkaline phosphatase (NEB) for 30 min. .. Pellets were suspended in 5 µl DNase I buffer and 0.2 U DNase I (NEB) and 0.2 U snake venom phosphodiesterase (Worthington) was added to the reactions.

Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
Article Snippet: .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion. ..

Activity Assay:

Article Title: A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA
Article Snippet: Paragraph title: β-gt specificity and activity assay ... Pellets were suspended in 5 µl DNase I buffer and 0.2 U DNase I (NEB) and 0.2 U snake venom phosphodiesterase (Worthington) was added to the reactions.

Expressing:

Article Title: A single catalytic domain of the junction-resolving enzyme T7 endonuclease I is a non-specific nicking endonuclease
Article Snippet: .. Materials Restriction enzymes, nicking enzyme N.BstNB I, DNA polymerases, T4 ligase, T4 DNA kinase, β-agarase, λ Exonuclease, DNase I, the maltose-binding protein (MBP) protein fusion expression and purification system including plasmid pMAL-c2x, the host E.coli strains TB1 and ER2566, Factor Xa protease, genenase, the cruciform structure-containing plasmid pUC(AT), 2 logs DNA ladder and synthetic oligonucleotides were obtained from New England Biolabs Inc. (NEB). .. Plasmid pNB1 was a gift from Rebecca Kucera (NEB).

Magnetic Resonance Imaging:

Article Title: Generation of a Novel Nucleic Acid-Based Reporter System To Detect Phenotypic Susceptibility to Antibiotics in Mycobacterium tuberculosis
Article Snippet: .. BssHII, NotI, XbaI, Antarctic phosphatase, MRI, avian myeloblastosis virus reverse transcriptase, DNase I, and T4 DNA ligase were obtained from New England Biolabs. .. Taq polymerase and deoxyribonucleotides were obtained from GenScript Corporation.

other:

Article Title: The Principal Role of Ku in Telomere Length Maintenance Is Promotion of Est1 Association with Telomeres
Article Snippet: Furthermore, the interaction between Ku and Est1 and Est2 was not mediated by DNA as treatment of lysates with DNase I had no impact on the co-immunoprecipitation efficiency ( and Figure S3 A).

Article Title: Regulation of the vapBC-1 Toxin-Antitoxin Locus in Nontypeable Haemophilus influenzae
Article Snippet: Reactions were quenched with 75 µL of DNase I stop solution (90% ethanol/220 mM sodium acetate/70 ng/µL yeast tRNA), mixed, and placed immediately in a dry ice/ethanol bath for 30 min.

Polymerase Chain Reaction:

Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
Article Snippet: Then, the PCR fragment was extracted and purified from 1.5% agarose gel in TAE buffer using QIAquick Gel Extraction Kit (QIAGEN). .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion.

Binding Assay:

Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
Article Snippet: .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion. ..

Fluorescence:

Article Title: In-Cell RNA Hydrolysis Assay: A Method for the Determination of the RNase Activity of Potential RNases
Article Snippet: .. The fluorescence intensity of RiboGreen was similar in the presence and absence of DNase I (Fig. f). .. DNA molecules packaged into chromatin in the nucleus would not be attacked by a protein with DNA- and RNA-hydrolyzing activities, such as 3D8 scFv antibody; therefore, the majority of RiboGreen fluorescence in this assay can be attributed to RNA, not to DNA.

Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
Article Snippet: Individual footprint reactions were initiated with a 22 μl binding reaction mixture that contained 12.5 mM HEPES (pH 7.8), 5 mM K-acetate (pH 8.0), 2.5 mM Mg-acetate, 1 mM CaCl2 , 12.5 μg/ml bovine serum albumin [ ], 0.3 mg/ml heparin, 200 nM of fluorescence-labeled DNA probe and various amounts of purified protein. .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion.

Labeling:

Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
Article Snippet: DNase I footprint assay Fluorescently labeled DNA probes containing either the pucB or crtI promoter regions were prepared by PCR amplification. .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion.

Purification:

Article Title: A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA
Article Snippet: Reactions were terminated by the addition of 10 µl Stop Buffer [20 µg Protenase K, 50 mM Tris–HCl (pH 8.0), 100 mM EDTA (pH 8.0), 2% (w/v) SDS] followed by incubation at 42°C for 60 min. Oligonucleotides were purified by phenol:CHCl3 :IAA extraction followed by an ethanol precipitation. .. Pellets were suspended in 5 µl DNase I buffer and 0.2 U DNase I (NEB) and 0.2 U snake venom phosphodiesterase (Worthington) was added to the reactions.

Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
Article Snippet: Individual footprint reactions were initiated with a 22 μl binding reaction mixture that contained 12.5 mM HEPES (pH 7.8), 5 mM K-acetate (pH 8.0), 2.5 mM Mg-acetate, 1 mM CaCl2 , 12.5 μg/ml bovine serum albumin [ ], 0.3 mg/ml heparin, 200 nM of fluorescence-labeled DNA probe and various amounts of purified protein. .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion.

Plasmid Preparation:

Article Title: Human U6 promoter drives stronger shRNA activity than its schistosome orthologue in Schistosoma mansoni and human fibrosarcoma cells
Article Snippet: In brief, GP2-293 cells were co-transfected with the MLV encoding plasmid ( ) along with pVSVG, which encodes vesicular stomatitis virus glycoprotein (the pseudotyping envelope) using liposomes (Lipofectamine 2000, Invitrogen). .. Culture supernatants were collected 48 hours later, filtered through 0.45 μm pore size membranes, the filtrate incubated with DNAse I (New England Biolabs, Ipswich, MA) to remove contaminating plasmids, and virions in culture supernatants recovered by centrifugation (Sorvall SS34 rotor, 50,000 g, 90 minutes, 4°C).

shRNA:

Article Title: Human U6 promoter drives stronger shRNA activity than its schistosome orthologue in Schistosoma mansoni and human fibrosarcoma cells
Article Snippet: Culture supernatants were collected 48 hours later, filtered through 0.45 μm pore size membranes, the filtrate incubated with DNAse I (New England Biolabs, Ipswich, MA) to remove contaminating plasmids, and virions in culture supernatants recovered by centrifugation (Sorvall SS34 rotor, 50,000 g, 90 minutes, 4°C). .. Pelleted virions were resuspended in schistosomule wash medium (above), virion titers estimated using quantitative PCR , and aliquots of virions stored at -80°C until needed. pLNHXΔD70 was employed to produce control virions without shRNA encoding cassettes.

Agarose Gel Electrophoresis:

Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
Article Snippet: Then, the PCR fragment was extracted and purified from 1.5% agarose gel in TAE buffer using QIAquick Gel Extraction Kit (QIAGEN). .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion.

Ethanol Precipitation:

Article Title: A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA
Article Snippet: DNA was cleaned by phenol:CHCl3 :IAA (isoamyl alcohol) extraction followed by an ethanol precipitation. .. Pellets were suspended in 5 µl DNase I buffer and 0.2 U DNase I (NEB) and 0.2 U snake venom phosphodiesterase (Worthington) was added to the reactions.

Gel Extraction:

Article Title: Evidence that Altered Cis Element Spacing Affects PpsR Mediated Redox Control of Photosynthesis Gene Expression in Rubrivivax gelatinosus
Article Snippet: Then, the PCR fragment was extracted and purified from 1.5% agarose gel in TAE buffer using QIAquick Gel Extraction Kit (QIAGEN). .. Initial binding reaction mixtures were incubated for 30 min at 22°C followed by a 15 min DNase I digestion that was initiated by adding 3 μl of DNase I (New England Biolabs) at an approximately 1:100 dilution (0.02 units/μl) in a footprint binding buffer, which gave partial probe digestion.

Variant Assay:

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