nebuffer 4  (New England Biolabs)


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    Name:
    NEBuffer 4
    Description:
    NEBuffer 4 5 0 ml
    Catalog Number:
    b7004s
    Price:
    24
    Size:
    5 0 ml
    Category:
    Buffers
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    Structured Review

    New England Biolabs nebuffer 4
    NEBuffer 4
    NEBuffer 4 5 0 ml
    https://www.bioz.com/result/nebuffer 4/product/New England Biolabs
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    nebuffer 4 - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Rethinking the Evolution of Single-Stranded RNA (ssRNA) Bacteriophages Based on Genomic Sequences and Characterizations of Two R-Plasmid-Dependent ssRNA Phages, C-1 and Hgal1
    Article Snippet: Paragraph title: Phage genome cDNA synthesis, cloning, and sequencing. ... The 50-μl reaction mixture was prepared with 5 μl of 10× NEBuffer 4 (New England BioLabs), 20 μl cDNA, 0.5 μl of 10 mM dATP, 0.5 μl terminal transferase, 5 μl of 2.5 mM CoCl2 , and 19 μl nuclease-free water.

    Amplification:

    Article Title: Duplex Proximity Sequencing (Pro-Seq): A method to improve DNA sequencing accuracy without the cost of molecular barcoding redundancy
    Article Snippet: Samples were then amplified on a BioRad T100 thermocycler with an initial denaturation at 98 °C for 30 s, 38 cycles at 98 °C for 10 s, 60 °C for 30 s, and 72 °C for 30 s followed by a final hold at 4 °C. .. Un-linked DNA was then digested enzymatically in a 50 μL reaction by mixing the eluted DNA from the previous step with 6.7 Units of T7 Exonuclease (NEB), 41.7 units of RecJf (NEB) and nuclease free water (IDT) in a 1x final concentration of NEBuffer 4 (NEB).

    Article Title: Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
    Article Snippet: .. Eliminate parental DNA on amplification products by digestion of methylated DNA with DpnI for 1h 30min at 37°C: mix in a centrifuge tube 17.5μl of PCR product with 2μl of 10x NEBuffer 4 and 0.5μl of DpnI (10U; New England Biolabs). .. Transform DpnI-treated PCR product: mix 2μl of DpnI-treated DNA with 45μl of XL10-Gold Ultracompetent E. coli (Stratagene) in pre-chilled 15 ml cell culture tube.

    Terminal Restriction Fragment Length Polymorphism:

    Article Title: Impact of elevated atmospheric O3 on the actinobacterial community structure and function in the rhizosphere of European beech (Fagus sylvatica L.)
    Article Snippet: Paragraph title: TRFLP analysis ... For the analysis of PKS type II genes, 100 ng of the PCR products were digested with 20 U Hha I (New England Biolabs, Frankfurt am Main, Germany) in a total of 20 μ L of 1x NEBuffer 4 (New England Biolabs, Frankfurt am Main, Germany) supplemented with 2 μ g BSA.

    Article Title: Impact of Lactobacillus reuteri colonization on gut microbiota, inflammation, and crying time in infant colic
    Article Snippet: T-RFLP fragments were generated using separate reactions for enzymes Alu I and Sau 96I (New England Biolabs, Ipswich, USA). .. In 25 µl reactions, approximately 50–100 ng of purified PCR product was digested with 5 U of enzyme and 1 X NEBuffer 4 (New England Biolabs) in a water bath (37 °C overnight).

    Microarray:

    Article Title: Nature and distribution of large sequence polymorphisms in Saccharomyces cerevisiae
    Article Snippet: Paragraph title: DNA extraction and microarray hybridization ... The DNA fragments were 3′ end-labeled by incubation with 1 nmol of biotin-11-ddATP (Perkin Elmer) and 20 U of terminal deoxynucleotidyl transferase (New England Biolabs) in 1× NEBuffer 4 (New England Biolabs) for 1 h at 37 °C, and the terminal transferase was subsequently heat inactivated at 75 °C for 25 min. Hybridization of the target DNA onto Affymetrix GeneChip® S. cerevisiae Tiling 1.0R Arrays followed standard Affymetrix protocols for hybridization, washing and staining , and the arrays were scanned using the Affymetrix scanner at 0.7 μm resolution.

    Incubation:

    Article Title: Impact of elevated atmospheric O3 on the actinobacterial community structure and function in the rhizosphere of European beech (Fagus sylvatica L.)
    Article Snippet: For the analysis of PKS type II genes, 100 ng of the PCR products were digested with 20 U Hha I (New England Biolabs, Frankfurt am Main, Germany) in a total of 20 μ L of 1x NEBuffer 4 (New England Biolabs, Frankfurt am Main, Germany) supplemented with 2 μ g BSA. .. For the analysis of PKS type II genes, 100 ng of the PCR products were digested with 20 U Hha I (New England Biolabs, Frankfurt am Main, Germany) in a total of 20 μ L of 1x NEBuffer 4 (New England Biolabs, Frankfurt am Main, Germany) supplemented with 2 μ g BSA.

    Article Title: Rethinking the Evolution of Single-Stranded RNA (ssRNA) Bacteriophages Based on Genomic Sequences and Characterizations of Two R-Plasmid-Dependent ssRNA Phages, C-1 and Hgal1
    Article Snippet: The 50-μl reaction mixture was prepared with 5 μl of 10× NEBuffer 4 (New England BioLabs), 20 μl cDNA, 0.5 μl of 10 mM dATP, 0.5 μl terminal transferase, 5 μl of 2.5 mM CoCl2 , and 19 μl nuclease-free water. .. The mixture was then incubated at 37°C for 30 min, followed by heat inactivation at 70°C for 10 min.

    Article Title: Self-replication of circular DNA by a self-encoded DNA polymerase through rolling-circle replication and recombination
    Article Snippet: .. Reaction mixtures were diluted 10-fold with deionized distilled water and mixed with 200 mU/μl exonuclease V (New England BioLabs), 1 mM ATP, and NEBuffer 4 (New England BioLabs, 50 mM potassium acetate, 20 mM Tris acetate, 10 mM magnesium acetate, 1 mM DTT, pH 7.9), and incubated at 37 °C for 4 h. After incubation, the mixtures were purified and concentrated 10-fold using a DNA column (PureLink PCR micro Kit). .. After the TTcDR reaction, the linear DNAs were degraded by the exonucleases as described above and subjected to 1% agarose gel electrophoresis with a control circular DNA.

    Article Title: Nature and distribution of large sequence polymorphisms in Saccharomyces cerevisiae
    Article Snippet: .. The DNA fragments were 3′ end-labeled by incubation with 1 nmol of biotin-11-ddATP (Perkin Elmer) and 20 U of terminal deoxynucleotidyl transferase (New England Biolabs) in 1× NEBuffer 4 (New England Biolabs) for 1 h at 37 °C, and the terminal transferase was subsequently heat inactivated at 75 °C for 25 min. Hybridization of the target DNA onto Affymetrix GeneChip® S. cerevisiae Tiling 1.0R Arrays followed standard Affymetrix protocols for hybridization, washing and staining , and the arrays were scanned using the Affymetrix scanner at 0.7 μm resolution. .. Average hybridization intensities were computed for each oligonucleotide feature with the GeneChip® Operating Software (Affymetrix) using the hybridization intensities of the 9 central pixels.

    Article Title: HEx: A heterologous expression platform for the discovery of fungal natural products
    Article Snippet: .. A volume of 22.5 μl of each prepared plasmid was combined with 1.5 μl of Exonuclease V (10 U/μl; NEB, M0345L), 3.0 μl of NEB Buffer 4 (NEB, B7004S), and 3.0 μl of ATP (10 mM; NEB, P0756S) followed by incubation at 37°C for 1 hour. .. Exonuclease reactions were quenched by the addition of 1 μl of EDTA (0.33 M) and incubation for 30 min at 70°C.

    Modification:

    Article Title: HEx: A heterologous expression platform for the discovery of fungal natural products
    Article Snippet: Each pellet in the deep-well block was resuspended in 250 μl of this modified P1 and incubated with shaking for 2 to 3 hours. .. A volume of 22.5 μl of each prepared plasmid was combined with 1.5 μl of Exonuclease V (10 U/μl; NEB, M0345L), 3.0 μl of NEB Buffer 4 (NEB, B7004S), and 3.0 μl of ATP (10 mM; NEB, P0756S) followed by incubation at 37°C for 1 hour.

    Hybridization:

    Article Title: Nature and distribution of large sequence polymorphisms in Saccharomyces cerevisiae
    Article Snippet: .. The DNA fragments were 3′ end-labeled by incubation with 1 nmol of biotin-11-ddATP (Perkin Elmer) and 20 U of terminal deoxynucleotidyl transferase (New England Biolabs) in 1× NEBuffer 4 (New England Biolabs) for 1 h at 37 °C, and the terminal transferase was subsequently heat inactivated at 75 °C for 25 min. Hybridization of the target DNA onto Affymetrix GeneChip® S. cerevisiae Tiling 1.0R Arrays followed standard Affymetrix protocols for hybridization, washing and staining , and the arrays were scanned using the Affymetrix scanner at 0.7 μm resolution. .. Average hybridization intensities were computed for each oligonucleotide feature with the GeneChip® Operating Software (Affymetrix) using the hybridization intensities of the 9 central pixels.

    Ligation:

    Article Title: Streptococcus pneumoniae Folate Biosynthesis Responds to Environmental CO2 Levels
    Article Snippet: Briefly, a 200-μl solution with 2 μg S. pneumoniae mutant library genomic DNA in NEBuffer 4 (New England BioLabs) with 5 μM S -adenosylmethionine was digested with 10 U MmeI (New England BioLabs) for 4 h at 37°C and dephosphorylated with 1 U calf intestine alkaline phosphatase (Invitrogen) for 30 min at 50°C. .. Ligation of 100 ng dephosphorylated MmeI restriction fragments with 2 pmol phosphorylated adapter was performed in the presence of T4 DNA ligase buffer with 2 U T4 DNA ligase (New England BioLabs) in a total volume of 20 μl for 1 h at 16°C.

    Cell Culture:

    Article Title: Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
    Article Snippet: Eliminate parental DNA on amplification products by digestion of methylated DNA with DpnI for 1h 30min at 37°C: mix in a centrifuge tube 17.5μl of PCR product with 2μl of 10x NEBuffer 4 and 0.5μl of DpnI (10U; New England Biolabs). .. Transform DpnI-treated PCR product: mix 2μl of DpnI-treated DNA with 45μl of XL10-Gold Ultracompetent E. coli (Stratagene) in pre-chilled 15 ml cell culture tube.

    Generated:

    Article Title: Duplex Proximity Sequencing (Pro-Seq): A method to improve DNA sequencing accuracy without the cost of molecular barcoding redundancy
    Article Snippet: Droplets were generated on the RainDrop Source instrument (RainDance Technologies) using ThunderBolts Open Source consumables (RainDance Technologies) as per manufacturer’s specifications. .. Un-linked DNA was then digested enzymatically in a 50 μL reaction by mixing the eluted DNA from the previous step with 6.7 Units of T7 Exonuclease (NEB), 41.7 units of RecJf (NEB) and nuclease free water (IDT) in a 1x final concentration of NEBuffer 4 (NEB).

    Article Title: Impact of Lactobacillus reuteri colonization on gut microbiota, inflammation, and crying time in infant colic
    Article Snippet: T-RFLP fragments were generated using separate reactions for enzymes Alu I and Sau 96I (New England Biolabs, Ipswich, USA). .. In 25 µl reactions, approximately 50–100 ng of purified PCR product was digested with 5 U of enzyme and 1 X NEBuffer 4 (New England Biolabs) in a water bath (37 °C overnight).

    Sequencing:

    Article Title: Rethinking the Evolution of Single-Stranded RNA (ssRNA) Bacteriophages Based on Genomic Sequences and Characterizations of Two R-Plasmid-Dependent ssRNA Phages, C-1 and Hgal1
    Article Snippet: Paragraph title: Phage genome cDNA synthesis, cloning, and sequencing. ... The 50-μl reaction mixture was prepared with 5 μl of 10× NEBuffer 4 (New England BioLabs), 20 μl cDNA, 0.5 μl of 10 mM dATP, 0.5 μl terminal transferase, 5 μl of 2.5 mM CoCl2 , and 19 μl nuclease-free water.

    Article Title: Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
    Article Snippet: Paragraph title: 1. Site-directed mutagenesis of highly GC-rich KISS1R gene sequence ... Eliminate parental DNA on amplification products by digestion of methylated DNA with DpnI for 1h 30min at 37°C: mix in a centrifuge tube 17.5μl of PCR product with 2μl of 10x NEBuffer 4 and 0.5μl of DpnI (10U; New England Biolabs).

    Article Title: HEx: A heterologous expression platform for the discovery of fungal natural products
    Article Snippet: Paragraph title: Preparation of DNA for direct sequencing of yeast plasmid DNA ... A volume of 22.5 μl of each prepared plasmid was combined with 1.5 μl of Exonuclease V (10 U/μl; NEB, M0345L), 3.0 μl of NEB Buffer 4 (NEB, B7004S), and 3.0 μl of ATP (10 mM; NEB, P0756S) followed by incubation at 37°C for 1 hour.

    DNA Extraction:

    Article Title: Nature and distribution of large sequence polymorphisms in Saccharomyces cerevisiae
    Article Snippet: Paragraph title: DNA extraction and microarray hybridization ... The DNA fragments were 3′ end-labeled by incubation with 1 nmol of biotin-11-ddATP (Perkin Elmer) and 20 U of terminal deoxynucleotidyl transferase (New England Biolabs) in 1× NEBuffer 4 (New England Biolabs) for 1 h at 37 °C, and the terminal transferase was subsequently heat inactivated at 75 °C for 25 min. Hybridization of the target DNA onto Affymetrix GeneChip® S. cerevisiae Tiling 1.0R Arrays followed standard Affymetrix protocols for hybridization, washing and staining , and the arrays were scanned using the Affymetrix scanner at 0.7 μm resolution.

    Methylation:

    Article Title: Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
    Article Snippet: .. Eliminate parental DNA on amplification products by digestion of methylated DNA with DpnI for 1h 30min at 37°C: mix in a centrifuge tube 17.5μl of PCR product with 2μl of 10x NEBuffer 4 and 0.5μl of DpnI (10U; New England Biolabs). .. Transform DpnI-treated PCR product: mix 2μl of DpnI-treated DNA with 45μl of XL10-Gold Ultracompetent E. coli (Stratagene) in pre-chilled 15 ml cell culture tube.

    Mutagenesis:

    Article Title: Streptococcus pneumoniae Folate Biosynthesis Responds to Environmental CO2 Levels
    Article Snippet: .. Briefly, a 200-μl solution with 2 μg S. pneumoniae mutant library genomic DNA in NEBuffer 4 (New England BioLabs) with 5 μM S -adenosylmethionine was digested with 10 U MmeI (New England BioLabs) for 4 h at 37°C and dephosphorylated with 1 U calf intestine alkaline phosphatase (Invitrogen) for 30 min at 50°C. ..

    Article Title: Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
    Article Snippet: Paragraph title: 1. Site-directed mutagenesis of highly GC-rich KISS1R gene sequence ... Eliminate parental DNA on amplification products by digestion of methylated DNA with DpnI for 1h 30min at 37°C: mix in a centrifuge tube 17.5μl of PCR product with 2μl of 10x NEBuffer 4 and 0.5μl of DpnI (10U; New England Biolabs).

    Isolation:

    Article Title: Rethinking the Evolution of Single-Stranded RNA (ssRNA) Bacteriophages Based on Genomic Sequences and Characterizations of Two R-Plasmid-Dependent ssRNA Phages, C-1 and Hgal1
    Article Snippet: RNA was isolated by phenol-chloroform extraction and ethanol precipitation. .. The 50-μl reaction mixture was prepared with 5 μl of 10× NEBuffer 4 (New England BioLabs), 20 μl cDNA, 0.5 μl of 10 mM dATP, 0.5 μl terminal transferase, 5 μl of 2.5 mM CoCl2 , and 19 μl nuclease-free water.

    Article Title: HEx: A heterologous expression platform for the discovery of fungal natural products
    Article Snippet: Plasmid DNA was isolated using a modified version of the QIAprep Turbo miniprep kit (27173/27191/27193, Qiagen). .. A volume of 22.5 μl of each prepared plasmid was combined with 1.5 μl of Exonuclease V (10 U/μl; NEB, M0345L), 3.0 μl of NEB Buffer 4 (NEB, B7004S), and 3.0 μl of ATP (10 mM; NEB, P0756S) followed by incubation at 37°C for 1 hour.

    Labeling:

    Article Title: Impact of elevated atmospheric O3 on the actinobacterial community structure and function in the rhizosphere of European beech (Fagus sylvatica L.)
    Article Snippet: For the analysis of PKS type II genes, 100 ng of the PCR products were digested with 20 U Hha I (New England Biolabs, Frankfurt am Main, Germany) in a total of 20 μ L of 1x NEBuffer 4 (New England Biolabs, Frankfurt am Main, Germany) supplemented with 2 μ g BSA. .. For detection of labeled fragments 2.5 μ L of the purified digestion reaction was mixed with 0.25 μ L GenomeLab DNA Size Standard 600 (Beckman Coulter GmbH, Krefeld, Germany) and 27.25 μ L SLS buffer (Beckman Coulter GmbH, Krefeld, Germany).

    Purification:

    Article Title: Impact of elevated atmospheric O3 on the actinobacterial community structure and function in the rhizosphere of European beech (Fagus sylvatica L.)
    Article Snippet: For the analysis of PKS type II genes, 100 ng of the PCR products were digested with 20 U Hha I (New England Biolabs, Frankfurt am Main, Germany) in a total of 20 μ L of 1x NEBuffer 4 (New England Biolabs, Frankfurt am Main, Germany) supplemented with 2 μ g BSA. .. For the analysis of PKS type II genes, 100 ng of the PCR products were digested with 20 U Hha I (New England Biolabs, Frankfurt am Main, Germany) in a total of 20 μ L of 1x NEBuffer 4 (New England Biolabs, Frankfurt am Main, Germany) supplemented with 2 μ g BSA.

    Article Title: Rethinking the Evolution of Single-Stranded RNA (ssRNA) Bacteriophages Based on Genomic Sequences and Characterizations of Two R-Plasmid-Dependent ssRNA Phages, C-1 and Hgal1
    Article Snippet: The 50-μl reaction mixture was prepared with 5 μl of 10× NEBuffer 4 (New England BioLabs), 20 μl cDNA, 0.5 μl of 10 mM dATP, 0.5 μl terminal transferase, 5 μl of 2.5 mM CoCl2 , and 19 μl nuclease-free water. .. The resulting product was purified (QIAquick PCR purification kit; Qiagen), end repaired (with T4 DNA polymerase; Fermentas), and then ligated (with T4 DNA ligase, Fermentas) to pUC19 cut with SmaI (Fermentas).

    Article Title: Self-replication of circular DNA by a self-encoded DNA polymerase through rolling-circle replication and recombination
    Article Snippet: .. Reaction mixtures were diluted 10-fold with deionized distilled water and mixed with 200 mU/μl exonuclease V (New England BioLabs), 1 mM ATP, and NEBuffer 4 (New England BioLabs, 50 mM potassium acetate, 20 mM Tris acetate, 10 mM magnesium acetate, 1 mM DTT, pH 7.9), and incubated at 37 °C for 4 h. After incubation, the mixtures were purified and concentrated 10-fold using a DNA column (PureLink PCR micro Kit). .. After the TTcDR reaction, the linear DNAs were degraded by the exonucleases as described above and subjected to 1% agarose gel electrophoresis with a control circular DNA.

    Article Title: Impact of Lactobacillus reuteri colonization on gut microbiota, inflammation, and crying time in infant colic
    Article Snippet: .. In 25 µl reactions, approximately 50–100 ng of purified PCR product was digested with 5 U of enzyme and 1 X NEBuffer 4 (New England Biolabs) in a water bath (37 °C overnight). .. PCR products were precipitated and analysed by an AB3730 DNA analyser using AB GeneMapper software (Applied Biosystems, Carlsbad, USA) at the Australian Genome Research Facility, Parkville, Australia.

    Article Title: HEx: A heterologous expression platform for the discovery of fungal natural products
    Article Snippet: A volume of 22.5 μl of each prepared plasmid was combined with 1.5 μl of Exonuclease V (10 U/μl; NEB, M0345L), 3.0 μl of NEB Buffer 4 (NEB, B7004S), and 3.0 μl of ATP (10 mM; NEB, P0756S) followed by incubation at 37°C for 1 hour. .. Finally, reactions were purified using Sera-mag magnetic particles (1.5 mg/ml, bead/sample ratio = 1:1) with final elution in 25 μl of tris-Cl (10 mM).

    Polymerase Chain Reaction:

    Article Title: Impact of elevated atmospheric O3 on the actinobacterial community structure and function in the rhizosphere of European beech (Fagus sylvatica L.)
    Article Snippet: .. For the analysis of PKS type II genes, 100 ng of the PCR products were digested with 20 U Hha I (New England Biolabs, Frankfurt am Main, Germany) in a total of 20 μ L of 1x NEBuffer 4 (New England Biolabs, Frankfurt am Main, Germany) supplemented with 2 μ g BSA. .. For the analysis of PKS type II genes, 100 ng of the PCR products were digested with 20 U Hha I (New England Biolabs, Frankfurt am Main, Germany) in a total of 20 μ L of 1x NEBuffer 4 (New England Biolabs, Frankfurt am Main, Germany) supplemented with 2 μ g BSA.

    Article Title: Rethinking the Evolution of Single-Stranded RNA (ssRNA) Bacteriophages Based on Genomic Sequences and Characterizations of Two R-Plasmid-Dependent ssRNA Phages, C-1 and Hgal1
    Article Snippet: The 50-μl reaction mixture was prepared with 5 μl of 10× NEBuffer 4 (New England BioLabs), 20 μl cDNA, 0.5 μl of 10 mM dATP, 0.5 μl terminal transferase, 5 μl of 2.5 mM CoCl2 , and 19 μl nuclease-free water. .. The resulting product was purified (QIAquick PCR purification kit; Qiagen), end repaired (with T4 DNA polymerase; Fermentas), and then ligated (with T4 DNA ligase, Fermentas) to pUC19 cut with SmaI (Fermentas).

    Article Title: Self-replication of circular DNA by a self-encoded DNA polymerase through rolling-circle replication and recombination
    Article Snippet: .. Reaction mixtures were diluted 10-fold with deionized distilled water and mixed with 200 mU/μl exonuclease V (New England BioLabs), 1 mM ATP, and NEBuffer 4 (New England BioLabs, 50 mM potassium acetate, 20 mM Tris acetate, 10 mM magnesium acetate, 1 mM DTT, pH 7.9), and incubated at 37 °C for 4 h. After incubation, the mixtures were purified and concentrated 10-fold using a DNA column (PureLink PCR micro Kit). .. After the TTcDR reaction, the linear DNAs were degraded by the exonucleases as described above and subjected to 1% agarose gel electrophoresis with a control circular DNA.

    Article Title: Impact of Lactobacillus reuteri colonization on gut microbiota, inflammation, and crying time in infant colic
    Article Snippet: .. In 25 µl reactions, approximately 50–100 ng of purified PCR product was digested with 5 U of enzyme and 1 X NEBuffer 4 (New England Biolabs) in a water bath (37 °C overnight). .. PCR products were precipitated and analysed by an AB3730 DNA analyser using AB GeneMapper software (Applied Biosystems, Carlsbad, USA) at the Australian Genome Research Facility, Parkville, Australia.

    Article Title: Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
    Article Snippet: .. Eliminate parental DNA on amplification products by digestion of methylated DNA with DpnI for 1h 30min at 37°C: mix in a centrifuge tube 17.5μl of PCR product with 2μl of 10x NEBuffer 4 and 0.5μl of DpnI (10U; New England Biolabs). .. Transform DpnI-treated PCR product: mix 2μl of DpnI-treated DNA with 45μl of XL10-Gold Ultracompetent E. coli (Stratagene) in pre-chilled 15 ml cell culture tube.

    Blocking Assay:

    Article Title: HEx: A heterologous expression platform for the discovery of fungal natural products
    Article Snippet: Each pellet in the deep-well block was resuspended in 250 μl of this modified P1 and incubated with shaking for 2 to 3 hours. .. A volume of 22.5 μl of each prepared plasmid was combined with 1.5 μl of Exonuclease V (10 U/μl; NEB, M0345L), 3.0 μl of NEB Buffer 4 (NEB, B7004S), and 3.0 μl of ATP (10 mM; NEB, P0756S) followed by incubation at 37°C for 1 hour.

    Plasmid Preparation:

    Article Title: Mutagenesis and Analysis of Genetic Mutations in the GC-rich KISS1 Receptor Sequence Identified in Humans with Reproductive Disorders
    Article Snippet: In summary: Both (forward and reverse) primers must contain the desired mutation and anneal to the same sequence on opposite strands of the plasmid (e.g. forward and reverse primers are complementary to each other) Primers should be 25 to 45 bases long and end in one or more C or G bases Introduced mutation(s) should be in the middle of primer and flanked by ˜10–15 bases of correct sequence on both sides Melting temperature (Tm) of primers should be equal or greater than 78°C. .. Eliminate parental DNA on amplification products by digestion of methylated DNA with DpnI for 1h 30min at 37°C: mix in a centrifuge tube 17.5μl of PCR product with 2μl of 10x NEBuffer 4 and 0.5μl of DpnI (10U; New England Biolabs).

    Article Title: HEx: A heterologous expression platform for the discovery of fungal natural products
    Article Snippet: .. A volume of 22.5 μl of each prepared plasmid was combined with 1.5 μl of Exonuclease V (10 U/μl; NEB, M0345L), 3.0 μl of NEB Buffer 4 (NEB, B7004S), and 3.0 μl of ATP (10 mM; NEB, P0756S) followed by incubation at 37°C for 1 hour. .. Exonuclease reactions were quenched by the addition of 1 μl of EDTA (0.33 M) and incubation for 30 min at 70°C.

    Software:

    Article Title: Nature and distribution of large sequence polymorphisms in Saccharomyces cerevisiae
    Article Snippet: The DNA fragments were 3′ end-labeled by incubation with 1 nmol of biotin-11-ddATP (Perkin Elmer) and 20 U of terminal deoxynucleotidyl transferase (New England Biolabs) in 1× NEBuffer 4 (New England Biolabs) for 1 h at 37 °C, and the terminal transferase was subsequently heat inactivated at 75 °C for 25 min. Hybridization of the target DNA onto Affymetrix GeneChip® S. cerevisiae Tiling 1.0R Arrays followed standard Affymetrix protocols for hybridization, washing and staining , and the arrays were scanned using the Affymetrix scanner at 0.7 μm resolution. .. Average hybridization intensities were computed for each oligonucleotide feature with the GeneChip® Operating Software (Affymetrix) using the hybridization intensities of the 9 central pixels.

    Article Title: Impact of Lactobacillus reuteri colonization on gut microbiota, inflammation, and crying time in infant colic
    Article Snippet: In 25 µl reactions, approximately 50–100 ng of purified PCR product was digested with 5 U of enzyme and 1 X NEBuffer 4 (New England Biolabs) in a water bath (37 °C overnight). .. PCR products were precipitated and analysed by an AB3730 DNA analyser using AB GeneMapper software (Applied Biosystems, Carlsbad, USA) at the Australian Genome Research Facility, Parkville, Australia.

    Agarose Gel Electrophoresis:

    Article Title: Nature and distribution of large sequence polymorphisms in Saccharomyces cerevisiae
    Article Snippet: After heat inactivation of the DNase I by incubation of the digestion mixture at 95 °C for 20 min, the digested DNA was analyzed on a 2% agarose gel. .. The DNA fragments were 3′ end-labeled by incubation with 1 nmol of biotin-11-ddATP (Perkin Elmer) and 20 U of terminal deoxynucleotidyl transferase (New England Biolabs) in 1× NEBuffer 4 (New England Biolabs) for 1 h at 37 °C, and the terminal transferase was subsequently heat inactivated at 75 °C for 25 min. Hybridization of the target DNA onto Affymetrix GeneChip® S. cerevisiae Tiling 1.0R Arrays followed standard Affymetrix protocols for hybridization, washing and staining , and the arrays were scanned using the Affymetrix scanner at 0.7 μm resolution.

    Ethanol Precipitation:

    Article Title: Rethinking the Evolution of Single-Stranded RNA (ssRNA) Bacteriophages Based on Genomic Sequences and Characterizations of Two R-Plasmid-Dependent ssRNA Phages, C-1 and Hgal1
    Article Snippet: RNA was isolated by phenol-chloroform extraction and ethanol precipitation. .. The 50-μl reaction mixture was prepared with 5 μl of 10× NEBuffer 4 (New England BioLabs), 20 μl cDNA, 0.5 μl of 10 mM dATP, 0.5 μl terminal transferase, 5 μl of 2.5 mM CoCl2 , and 19 μl nuclease-free water.

    Concentration Assay:

    Article Title: Duplex Proximity Sequencing (Pro-Seq): A method to improve DNA sequencing accuracy without the cost of molecular barcoding redundancy
    Article Snippet: .. Un-linked DNA was then digested enzymatically in a 50 μL reaction by mixing the eluted DNA from the previous step with 6.7 Units of T7 Exonuclease (NEB), 41.7 units of RecJf (NEB) and nuclease free water (IDT) in a 1x final concentration of NEBuffer 4 (NEB). ..

    Staining:

    Article Title: Nature and distribution of large sequence polymorphisms in Saccharomyces cerevisiae
    Article Snippet: .. The DNA fragments were 3′ end-labeled by incubation with 1 nmol of biotin-11-ddATP (Perkin Elmer) and 20 U of terminal deoxynucleotidyl transferase (New England Biolabs) in 1× NEBuffer 4 (New England Biolabs) for 1 h at 37 °C, and the terminal transferase was subsequently heat inactivated at 75 °C for 25 min. Hybridization of the target DNA onto Affymetrix GeneChip® S. cerevisiae Tiling 1.0R Arrays followed standard Affymetrix protocols for hybridization, washing and staining , and the arrays were scanned using the Affymetrix scanner at 0.7 μm resolution. .. Average hybridization intensities were computed for each oligonucleotide feature with the GeneChip® Operating Software (Affymetrix) using the hybridization intensities of the 9 central pixels.

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    New England Biolabs nebuffer 4
    Nebuffer 4, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nebuffer 4/product/New England Biolabs
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