r3150  (New England Biolabs)


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    Name:
    PvuI HF
    Description:
    PvuI HF 2 500 units
    Catalog Number:
    R3150L
    Price:
    298
    Category:
    Restriction Enzymes
    Size:
    2 500 units
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    Structured Review

    New England Biolabs r3150
    PvuI HF
    PvuI HF 2 500 units
    https://www.bioz.com/result/r3150/product/New England Biolabs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    r3150 - by Bioz Stars, 2021-06
    93/100 stars

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    Related Articles

    Transfection:

    Article Title: Efficient derivation of extended pluripotent stem cells from NOD-scid Il2rg−/− mice
    Article Snippet: The expression cassette containing Ef1α promoter-Tdtomato-SV40 polyA-LoxP-PGK promoter-Puro-bGH polyA-LoxP was constructed into pEASY-Blunt vector (TransGen Biotech, CB501) for targeting Rosa26 locus. .. Before transfection, the targeting vector was linearized by PvuI (New England BioLabs, R3150L). .. A total of 2 × 106 cells were mixed with 10 μg of modified pX330 plasmid and 10 μg of targeting vector in final volume of 100 μL containing 82 μL P3 primary cell solution and 18 μL supplement 1 (PBP3-02250).

    Plasmid Preparation:

    Article Title: Efficient derivation of extended pluripotent stem cells from NOD-scid Il2rg−/− mice
    Article Snippet: The expression cassette containing Ef1α promoter-Tdtomato-SV40 polyA-LoxP-PGK promoter-Puro-bGH polyA-LoxP was constructed into pEASY-Blunt vector (TransGen Biotech, CB501) for targeting Rosa26 locus. .. Before transfection, the targeting vector was linearized by PvuI (New England BioLabs, R3150L). .. A total of 2 × 106 cells were mixed with 10 μg of modified pX330 plasmid and 10 μg of targeting vector in final volume of 100 μL containing 82 μL P3 primary cell solution and 18 μL supplement 1 (PBP3-02250).

    Article Title: A Targeted Vaccine against COVID-19: S1-Fc Vaccine Targeting the Antigen-Presenting Cell Compartment Elicits Protection against SARS-CoV-2 Infection
    Article Snippet: .. Production of linearized S1-Fc-encoding S1-Fc dsDNA fragmentThe plasmid pAAV S1-Fc was digested with New England Biolabs (NEB) Restriction Enzymes Pvu I-HF (Cat. No. R3150), Sph I-HF (Cat. No. R3182) and Mlu I-HF (Cat. No. R3198) at 37°C for 2.5 hours. .. The resulted fragments were separated by 1% agarose DNA gel electrophoresis.

    Polymerase Chain Reaction:

    Article Title: Expression of GBGT1 is epigenetically regulated by DNA methylation in ovarian cancer cells
    Article Snippet: .. Restriction fragment length analysis was performed on PCR products incubated with the restriction endonucleases AciI (methylated 50 bp + 167 bp, unmethylated 217 bp), PvuI-HF (methylated 37 bp + 180 bp, unmethylated 217 bp) and BsaAI (methylated 192 bp + 25 bp, unmethylated 217 bp) (New England BioLabs Inc., Genesearch Pty Ltd., Arundal, Queensland, Australia). .. Genomic DNA isolated from human blood serum served as an unmethylated COBRA control.

    Incubation:

    Article Title: Expression of GBGT1 is epigenetically regulated by DNA methylation in ovarian cancer cells
    Article Snippet: .. Restriction fragment length analysis was performed on PCR products incubated with the restriction endonucleases AciI (methylated 50 bp + 167 bp, unmethylated 217 bp), PvuI-HF (methylated 37 bp + 180 bp, unmethylated 217 bp) and BsaAI (methylated 192 bp + 25 bp, unmethylated 217 bp) (New England BioLabs Inc., Genesearch Pty Ltd., Arundal, Queensland, Australia). .. Genomic DNA isolated from human blood serum served as an unmethylated COBRA control.

    Article Title: Single-cell RNA-seq analysis reveals ploidy-dependent and cell-specific transcriptome changes in Arabidopsis female gametophytes
    Article Snippet: For library construction, 200 pg amplified DNA was tagmented using Nextera XT DNA sample preparation kit according to the manufacturer’s instructions (Illumina, San Diego, CA). .. The fragmented DNA was purified using AMPure XP beads (1:1 ratio; Beckman Coulter) and incubated with PvuI-HF enzyme (NEB, Ipswich, MA) for 30 min at 37 °C. .. After digestion by PvuI-HF, DNA was purified using AMPure XP beads (1:1 ratio; Beckman Coulter) and suspended in 18 μl water.

    Methylation:

    Article Title: Expression of GBGT1 is epigenetically regulated by DNA methylation in ovarian cancer cells
    Article Snippet: .. Restriction fragment length analysis was performed on PCR products incubated with the restriction endonucleases AciI (methylated 50 bp + 167 bp, unmethylated 217 bp), PvuI-HF (methylated 37 bp + 180 bp, unmethylated 217 bp) and BsaAI (methylated 192 bp + 25 bp, unmethylated 217 bp) (New England BioLabs Inc., Genesearch Pty Ltd., Arundal, Queensland, Australia). .. Genomic DNA isolated from human blood serum served as an unmethylated COBRA control.

    In Vitro:

    Article Title: hnRNPK Recruits PCGF3/5-PRC1 to the Xist RNA B-Repeat to Establish Polycomb-Mediated Chromosomal Silencing
    Article Snippet: 1 μL of 4sU-labeled RNA was quality-checked using the Agilent RNA 6000 Pico kit (Agilent technologies) according to the manufacturer’s instructions, and run on a 2100 Bioanalyzer Instrument (Agilent). .. Xist in vitro transcription RNAs were transcribed from a T7 promoter, following linearization with excess of PvuI-HF (NEB). .. 1 μg of DNA was in vitro transcribed using the HiScribe T7 High Yield RNA Synthesis Kit (NEB), according to the manufacturer’s guidelines.

    Purification:

    Article Title: Single-cell RNA-seq analysis reveals ploidy-dependent and cell-specific transcriptome changes in Arabidopsis female gametophytes
    Article Snippet: For library construction, 200 pg amplified DNA was tagmented using Nextera XT DNA sample preparation kit according to the manufacturer’s instructions (Illumina, San Diego, CA). .. The fragmented DNA was purified using AMPure XP beads (1:1 ratio; Beckman Coulter) and incubated with PvuI-HF enzyme (NEB, Ipswich, MA) for 30 min at 37 °C. .. After digestion by PvuI-HF, DNA was purified using AMPure XP beads (1:1 ratio; Beckman Coulter) and suspended in 18 μl water.

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  • 93
    New England Biolabs pvui hf
    DNA methylation of GBGT1 promoter region. (A) Illustration of the organization of the CpG island encompassing the transcription start site of the GBGT1 gene. The horizontal black line represents the 217 bp amplicon generated using COBRA and bisulfite sequencing. Vertical lines represent the organization of individual CpG dinucleotides within the amplified region. Black triangles indicate the respective recognition sites of the restriction endonucleases ( Acil, Pvul and BsaAl ). Each of these enzymes digested the amplicon only when the respective CpG dinucleotide(s) within the enzyme recognition site was methylated in the original DNA prior to bisulfite conversion ( <t>AciI</t> -active 50 + 176 bp; <t>PvuI</t> -active 37 + 180 bp; BsaAI -active 192 + 25 bp). (B) COBRA with each of the three different endonucleases ( Acil, Pvul, BsaAl ) revealed considerable variation in the methylation status among the cell lines. The degree of methylation (which relates to the intensity of the lower digested bands as compared to the upper undigested band) was consistent between each restriction enzyme for each cell line. (C) Methylation profiles of individual CpG sites from single DNA strands derived from bisulfite sequencing in A2780, HOSE17-1, OVCAR3, and SKOV3. Columns represent individual CpG sites. Rows represent number of sequenced clones (n = 12). Methylated CpG (black), unmethylated (grey), unknown status (white). (D) Restoration of GBGT1 expression in A2780 induced by treatment with 2.5 μM 5-Aza. RT-qPCR shows a time-dependent increase in GBGT1 transcription. Left, data are presented as the number of PCR products in 5-Aza treated samples relative to the mock-treated control (y-axis) as a function of time (24 h, 48 h, 72 h) after treatment (x-axis). Right, RT-qPCR products of GBGT1 and normalization control YWHAZ . (E) Western blot (autoradiograph and corresponding quantitative analysis) showing 5-Aza -induced increase in GBGT1 protein expression as a function of time after treatment in A2780.
    Pvui Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pvui hf/product/New England Biolabs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pvui hf - by Bioz Stars, 2021-06
    93/100 stars
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    DNA methylation of GBGT1 promoter region. (A) Illustration of the organization of the CpG island encompassing the transcription start site of the GBGT1 gene. The horizontal black line represents the 217 bp amplicon generated using COBRA and bisulfite sequencing. Vertical lines represent the organization of individual CpG dinucleotides within the amplified region. Black triangles indicate the respective recognition sites of the restriction endonucleases ( Acil, Pvul and BsaAl ). Each of these enzymes digested the amplicon only when the respective CpG dinucleotide(s) within the enzyme recognition site was methylated in the original DNA prior to bisulfite conversion ( AciI -active 50 + 176 bp; PvuI -active 37 + 180 bp; BsaAI -active 192 + 25 bp). (B) COBRA with each of the three different endonucleases ( Acil, Pvul, BsaAl ) revealed considerable variation in the methylation status among the cell lines. The degree of methylation (which relates to the intensity of the lower digested bands as compared to the upper undigested band) was consistent between each restriction enzyme for each cell line. (C) Methylation profiles of individual CpG sites from single DNA strands derived from bisulfite sequencing in A2780, HOSE17-1, OVCAR3, and SKOV3. Columns represent individual CpG sites. Rows represent number of sequenced clones (n = 12). Methylated CpG (black), unmethylated (grey), unknown status (white). (D) Restoration of GBGT1 expression in A2780 induced by treatment with 2.5 μM 5-Aza. RT-qPCR shows a time-dependent increase in GBGT1 transcription. Left, data are presented as the number of PCR products in 5-Aza treated samples relative to the mock-treated control (y-axis) as a function of time (24 h, 48 h, 72 h) after treatment (x-axis). Right, RT-qPCR products of GBGT1 and normalization control YWHAZ . (E) Western blot (autoradiograph and corresponding quantitative analysis) showing 5-Aza -induced increase in GBGT1 protein expression as a function of time after treatment in A2780.

    Journal: BMC Molecular Biology

    Article Title: Expression of GBGT1 is epigenetically regulated by DNA methylation in ovarian cancer cells

    doi: 10.1186/1471-2199-15-24

    Figure Lengend Snippet: DNA methylation of GBGT1 promoter region. (A) Illustration of the organization of the CpG island encompassing the transcription start site of the GBGT1 gene. The horizontal black line represents the 217 bp amplicon generated using COBRA and bisulfite sequencing. Vertical lines represent the organization of individual CpG dinucleotides within the amplified region. Black triangles indicate the respective recognition sites of the restriction endonucleases ( Acil, Pvul and BsaAl ). Each of these enzymes digested the amplicon only when the respective CpG dinucleotide(s) within the enzyme recognition site was methylated in the original DNA prior to bisulfite conversion ( AciI -active 50 + 176 bp; PvuI -active 37 + 180 bp; BsaAI -active 192 + 25 bp). (B) COBRA with each of the three different endonucleases ( Acil, Pvul, BsaAl ) revealed considerable variation in the methylation status among the cell lines. The degree of methylation (which relates to the intensity of the lower digested bands as compared to the upper undigested band) was consistent between each restriction enzyme for each cell line. (C) Methylation profiles of individual CpG sites from single DNA strands derived from bisulfite sequencing in A2780, HOSE17-1, OVCAR3, and SKOV3. Columns represent individual CpG sites. Rows represent number of sequenced clones (n = 12). Methylated CpG (black), unmethylated (grey), unknown status (white). (D) Restoration of GBGT1 expression in A2780 induced by treatment with 2.5 μM 5-Aza. RT-qPCR shows a time-dependent increase in GBGT1 transcription. Left, data are presented as the number of PCR products in 5-Aza treated samples relative to the mock-treated control (y-axis) as a function of time (24 h, 48 h, 72 h) after treatment (x-axis). Right, RT-qPCR products of GBGT1 and normalization control YWHAZ . (E) Western blot (autoradiograph and corresponding quantitative analysis) showing 5-Aza -induced increase in GBGT1 protein expression as a function of time after treatment in A2780.

    Article Snippet: Restriction fragment length analysis was performed on PCR products incubated with the restriction endonucleases AciI (methylated 50 bp + 167 bp, unmethylated 217 bp), PvuI-HF (methylated 37 bp + 180 bp, unmethylated 217 bp) and BsaAI (methylated 192 bp + 25 bp, unmethylated 217 bp) (New England BioLabs Inc., Genesearch Pty Ltd., Arundal, Queensland, Australia).

    Techniques: DNA Methylation Assay, Amplification, Generated, Combined Bisulfite Restriction Analysis Assay, Methylation Sequencing, Methylation, Derivative Assay, Clone Assay, Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Western Blot, Autoradiography