superhelical puc19  (New England Biolabs)


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  • 99
    Name:
    pUC19 Vector
    Description:
    pUC19 Vector 250 ug
    Catalog Number:
    N3041L
    Price:
    300
    Category:
    Vectors Plasmids
    Size:
    250 ug
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    Structured Review

    New England Biolabs superhelical puc19
    pUC19 Vector
    pUC19 Vector 250 ug
    https://www.bioz.com/result/superhelical puc19/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    superhelical puc19 - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "DNA oxidation profiles of copper phenanthrene chemical nucleases"

    Article Title: DNA oxidation profiles of copper phenanthrene chemical nucleases

    Journal: Frontiers in Chemistry

    doi: 10.3389/fchem.2015.00028

    Lane 1–4 (A–D) DNA cleavage reactions with 250 nM, 500 nM, 1.0 μM, and 2.5 μM test complex (A) Cu-Phen, (B) Cu-DPQ-Phen, (C) Cu-DPPZ-Phen, and (D) Cu-Terph, 400 ng superhelical pUC19 and 1 mM added Na-L-ascorbate incubated at 37°C for 30 min . Lanes 5–16 (A–D) DNA cleavage reactions in the presence of recognition elements, methyl green (MG), netropsin (Net), and [Co(NH 3 ) 6 ]Cl 3 (Co(III)), where 400 ng pUC19 was initially pre-treated with 8 μM of respective non-covalent binding control at 37° C for 45 min and then with 250 nM, 500 nM, 1 μM, and 2.5 μM test complex in the presence of 1 mM added Na-L-ascorbate at 37°C for 30 min.
    Figure Legend Snippet: Lane 1–4 (A–D) DNA cleavage reactions with 250 nM, 500 nM, 1.0 μM, and 2.5 μM test complex (A) Cu-Phen, (B) Cu-DPQ-Phen, (C) Cu-DPPZ-Phen, and (D) Cu-Terph, 400 ng superhelical pUC19 and 1 mM added Na-L-ascorbate incubated at 37°C for 30 min . Lanes 5–16 (A–D) DNA cleavage reactions in the presence of recognition elements, methyl green (MG), netropsin (Net), and [Co(NH 3 ) 6 ]Cl 3 (Co(III)), where 400 ng pUC19 was initially pre-treated with 8 μM of respective non-covalent binding control at 37° C for 45 min and then with 250 nM, 500 nM, 1 μM, and 2.5 μM test complex in the presence of 1 mM added Na-L-ascorbate at 37°C for 30 min.

    Techniques Used: Incubation, Binding Assay

    Related Articles

    Mutagenesis:

    Article Title: Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974 ▿
    Article Snippet: Genetic complementation was used to test the in vivo activities of SerRS1 and SerRS2. .. The host strain was an E. coli mutant (K28) that is ineffective at making its own serS gene product at high temperature ( , ). serS1 and serS2 were separately cloned into a high-copy vector, pUC19, under the control of a constitutive glnS ′ promoter, which is a glnS mutant promoter used for aaRS expression in E. coli , and the resulting plasmids were transformed into E. coli K28. ..

    Clone Assay:

    Article Title: Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974 ▿
    Article Snippet: Genetic complementation was used to test the in vivo activities of SerRS1 and SerRS2. .. The host strain was an E. coli mutant (K28) that is ineffective at making its own serS gene product at high temperature ( , ). serS1 and serS2 were separately cloned into a high-copy vector, pUC19, under the control of a constitutive glnS ′ promoter, which is a glnS mutant promoter used for aaRS expression in E. coli , and the resulting plasmids were transformed into E. coli K28. ..

    Article Title: Nucleosome Spacing Generated by ISWI and CHD1 Remodelers Is Constant Regardless of Nucleosome Density
    Article Snippet: .. Plasmids were isolated from Escherichia coli using the PureYield Maxiprep system (Promega). pUC19-PHO8 contains ∼3.5 kb of the Saccharomyces cerevisiae PHO8 locus cloned into pUC19 via BamHI and PstI, is 6,168 bp long, and is described as “pUC19-PHO8-long” in reference . pUC19-GCY1 contains ∼3.5 kb of the S. cerevisiae GCY1 locus (PCR product using primers 5′-CAGTCGGATGGAGCTCACTTCTATTGGCTTAGGAGC-3′ and 5′-CACTGTGCATTTCTAGAACGACGAAGACGAGGATTAG-3′ and genomic DNA of strain BY4741 [EUROSCARF] as the template) cloned into pUC19 via SacI and XbaI. pUC19-PHO8 and pUC19-GCY1 were linearized with BamHI (NEB), which cleaves right at or 400 bp downstream of, respectively, the upstream border between prokaryotic and eukaryotic DNA. .. The complete plasmids were used as the template for salt gradient dialysis (SGD) reconstitution, and complete linearization was confirmed by agarose gel electrophoresis prior to chromatin assembly.

    Article Title: Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974 ▿
    Article Snippet: The E. coli serS gene, including its native promoter and terminator, was amplified from E. coli JM109 genomic DNA using primers EcserS-F and EcserS-R. .. The product was cloned into the XbaI and EcoRI sites of pUC19 to produce pUC-EcSerRS. ..

    Plasmid Preparation:

    Article Title: Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974 ▿
    Article Snippet: Genetic complementation was used to test the in vivo activities of SerRS1 and SerRS2. .. The host strain was an E. coli mutant (K28) that is ineffective at making its own serS gene product at high temperature ( , ). serS1 and serS2 were separately cloned into a high-copy vector, pUC19, under the control of a constitutive glnS ′ promoter, which is a glnS mutant promoter used for aaRS expression in E. coli , and the resulting plasmids were transformed into E. coli K28. ..

    Article Title: Base-resolution detection of N4-methylcytosine in genomic DNA using 4mC-Tet-assisted-bisulfite- sequencing
    Article Snippet: Genomic DNA concentrations were determined using the Qubit® 2.0 fluorometer (Invitrogen) and the quality of DNA was assessed by agarose gel electrophoresis. .. Preparation of 304 bp model DNA with 4mC modifications For N 4 -methylcytosine (4mC) containing model DNA, 0.5 ng of pUC19 vector DNA (NEB) was PCR amplified as follows in a 50 μl reaction: 2.5 U RedTaq polymerase (Sigma), 5 μl 10× reaction buffer, 1 μl N 4 -methyl-dCTP (4mdCTP) (Trilink)/dATP/dGTP/dTTP cocktail (10 mM each), 1 μl 10 mM forward primer (5′-GAACGAAAACTCACGTTAAGGG), 1 μl 10 mM reverse primer (5′-TGCTGATAAATCTGGAGCCG). ..

    Article Title: C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents
    Article Snippet: Since condensation by C 3 opioids had been established by viscosity and turbidity measurements, visualization of DNA compaction was then followed by electrophoresis. .. Samples were titrated against both supercoiled pUC19 plasmid DNA and a 742 bp dsDNA fragment amplified from pUC19 encompassing the lacZα gene, before incubation for 5 h prior to analysis by agarose gel electrophoresis (Figure ). ..

    Expressing:

    Article Title: Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974 ▿
    Article Snippet: Genetic complementation was used to test the in vivo activities of SerRS1 and SerRS2. .. The host strain was an E. coli mutant (K28) that is ineffective at making its own serS gene product at high temperature ( , ). serS1 and serS2 were separately cloned into a high-copy vector, pUC19, under the control of a constitutive glnS ′ promoter, which is a glnS mutant promoter used for aaRS expression in E. coli , and the resulting plasmids were transformed into E. coli K28. ..

    Transformation Assay:

    Article Title: Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974 ▿
    Article Snippet: Genetic complementation was used to test the in vivo activities of SerRS1 and SerRS2. .. The host strain was an E. coli mutant (K28) that is ineffective at making its own serS gene product at high temperature ( , ). serS1 and serS2 were separately cloned into a high-copy vector, pUC19, under the control of a constitutive glnS ′ promoter, which is a glnS mutant promoter used for aaRS expression in E. coli , and the resulting plasmids were transformed into E. coli K28. ..

    Article Title: Secretory expression of biologically active human Herpes virus interleukin-10 analogues in Escherichia coli via a modified Sec-dependent transporter construct
    Article Snippet: After 4 h of incubation at 37°C, the amount of TNF-α released in the medium was measured in duplicate by ELISA (R & D Systems) according to the manufacturer’s instructions. .. Periplasmic fraction of pUC19 transformed E. coli BL21 (DE3) was used as TNF-α induction control. .. Commercial EBV IL-10 (R & D Systems) served as positive control.

    Isolation:

    Article Title: Nucleosome Spacing Generated by ISWI and CHD1 Remodelers Is Constant Regardless of Nucleosome Density
    Article Snippet: .. Plasmids were isolated from Escherichia coli using the PureYield Maxiprep system (Promega). pUC19-PHO8 contains ∼3.5 kb of the Saccharomyces cerevisiae PHO8 locus cloned into pUC19 via BamHI and PstI, is 6,168 bp long, and is described as “pUC19-PHO8-long” in reference . pUC19-GCY1 contains ∼3.5 kb of the S. cerevisiae GCY1 locus (PCR product using primers 5′-CAGTCGGATGGAGCTCACTTCTATTGGCTTAGGAGC-3′ and 5′-CACTGTGCATTTCTAGAACGACGAAGACGAGGATTAG-3′ and genomic DNA of strain BY4741 [EUROSCARF] as the template) cloned into pUC19 via SacI and XbaI. pUC19-PHO8 and pUC19-GCY1 were linearized with BamHI (NEB), which cleaves right at or 400 bp downstream of, respectively, the upstream border between prokaryotic and eukaryotic DNA. .. The complete plasmids were used as the template for salt gradient dialysis (SGD) reconstitution, and complete linearization was confirmed by agarose gel electrophoresis prior to chromatin assembly.

    Polymerase Chain Reaction:

    Article Title: Nucleosome Spacing Generated by ISWI and CHD1 Remodelers Is Constant Regardless of Nucleosome Density
    Article Snippet: .. Plasmids were isolated from Escherichia coli using the PureYield Maxiprep system (Promega). pUC19-PHO8 contains ∼3.5 kb of the Saccharomyces cerevisiae PHO8 locus cloned into pUC19 via BamHI and PstI, is 6,168 bp long, and is described as “pUC19-PHO8-long” in reference . pUC19-GCY1 contains ∼3.5 kb of the S. cerevisiae GCY1 locus (PCR product using primers 5′-CAGTCGGATGGAGCTCACTTCTATTGGCTTAGGAGC-3′ and 5′-CACTGTGCATTTCTAGAACGACGAAGACGAGGATTAG-3′ and genomic DNA of strain BY4741 [EUROSCARF] as the template) cloned into pUC19 via SacI and XbaI. pUC19-PHO8 and pUC19-GCY1 were linearized with BamHI (NEB), which cleaves right at or 400 bp downstream of, respectively, the upstream border between prokaryotic and eukaryotic DNA. .. The complete plasmids were used as the template for salt gradient dialysis (SGD) reconstitution, and complete linearization was confirmed by agarose gel electrophoresis prior to chromatin assembly.

    Article Title: Base-resolution detection of N4-methylcytosine in genomic DNA using 4mC-Tet-assisted-bisulfite- sequencing
    Article Snippet: Genomic DNA concentrations were determined using the Qubit® 2.0 fluorometer (Invitrogen) and the quality of DNA was assessed by agarose gel electrophoresis. .. Preparation of 304 bp model DNA with 4mC modifications For N 4 -methylcytosine (4mC) containing model DNA, 0.5 ng of pUC19 vector DNA (NEB) was PCR amplified as follows in a 50 μl reaction: 2.5 U RedTaq polymerase (Sigma), 5 μl 10× reaction buffer, 1 μl N 4 -methyl-dCTP (4mdCTP) (Trilink)/dATP/dGTP/dTTP cocktail (10 mM each), 1 μl 10 mM forward primer (5′-GAACGAAAACTCACGTTAAGGG), 1 μl 10 mM reverse primer (5′-TGCTGATAAATCTGGAGCCG). ..

    Amplification:

    Article Title: Base-resolution detection of N4-methylcytosine in genomic DNA using 4mC-Tet-assisted-bisulfite- sequencing
    Article Snippet: Genomic DNA concentrations were determined using the Qubit® 2.0 fluorometer (Invitrogen) and the quality of DNA was assessed by agarose gel electrophoresis. .. Preparation of 304 bp model DNA with 4mC modifications For N 4 -methylcytosine (4mC) containing model DNA, 0.5 ng of pUC19 vector DNA (NEB) was PCR amplified as follows in a 50 μl reaction: 2.5 U RedTaq polymerase (Sigma), 5 μl 10× reaction buffer, 1 μl N 4 -methyl-dCTP (4mdCTP) (Trilink)/dATP/dGTP/dTTP cocktail (10 mM each), 1 μl 10 mM forward primer (5′-GAACGAAAACTCACGTTAAGGG), 1 μl 10 mM reverse primer (5′-TGCTGATAAATCTGGAGCCG). ..

    Article Title: C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents
    Article Snippet: Since condensation by C 3 opioids had been established by viscosity and turbidity measurements, visualization of DNA compaction was then followed by electrophoresis. .. Samples were titrated against both supercoiled pUC19 plasmid DNA and a 742 bp dsDNA fragment amplified from pUC19 encompassing the lacZα gene, before incubation for 5 h prior to analysis by agarose gel electrophoresis (Figure ). ..

    Incubation:

    Article Title: C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents
    Article Snippet: Since condensation by C 3 opioids had been established by viscosity and turbidity measurements, visualization of DNA compaction was then followed by electrophoresis. .. Samples were titrated against both supercoiled pUC19 plasmid DNA and a 742 bp dsDNA fragment amplified from pUC19 encompassing the lacZα gene, before incubation for 5 h prior to analysis by agarose gel electrophoresis (Figure ). ..

    Agarose Gel Electrophoresis:

    Article Title: C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents
    Article Snippet: Since condensation by C 3 opioids had been established by viscosity and turbidity measurements, visualization of DNA compaction was then followed by electrophoresis. .. Samples were titrated against both supercoiled pUC19 plasmid DNA and a 742 bp dsDNA fragment amplified from pUC19 encompassing the lacZα gene, before incubation for 5 h prior to analysis by agarose gel electrophoresis (Figure ). ..

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  • 99
    New England Biolabs superhelical puc19
    Lane 1–4 (A–D) DNA cleavage reactions with 250 nM, 500 nM, 1.0 μM, and 2.5 μM test complex (A) Cu-Phen, (B) Cu-DPQ-Phen, (C) Cu-DPPZ-Phen, and (D) Cu-Terph, 400 ng <t>superhelical</t> <t>pUC19</t> and 1 mM added Na-L-ascorbate incubated at 37°C for 30 min . Lanes 5–16 (A–D) DNA cleavage reactions in the presence of recognition elements, methyl green (MG), netropsin (Net), and [Co(NH 3 ) 6 ]Cl 3 (Co(III)), where 400 ng pUC19 was initially pre-treated with 8 μM of respective non-covalent binding control at 37° C for 45 min and then with 250 nM, 500 nM, 1 μM, and 2.5 μM test complex in the presence of 1 mM added Na-L-ascorbate at 37°C for 30 min.
    Superhelical Puc19, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superhelical puc19/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    superhelical puc19 - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

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    Lane 1–4 (A–D) DNA cleavage reactions with 250 nM, 500 nM, 1.0 μM, and 2.5 μM test complex (A) Cu-Phen, (B) Cu-DPQ-Phen, (C) Cu-DPPZ-Phen, and (D) Cu-Terph, 400 ng superhelical pUC19 and 1 mM added Na-L-ascorbate incubated at 37°C for 30 min . Lanes 5–16 (A–D) DNA cleavage reactions in the presence of recognition elements, methyl green (MG), netropsin (Net), and [Co(NH 3 ) 6 ]Cl 3 (Co(III)), where 400 ng pUC19 was initially pre-treated with 8 μM of respective non-covalent binding control at 37° C for 45 min and then with 250 nM, 500 nM, 1 μM, and 2.5 μM test complex in the presence of 1 mM added Na-L-ascorbate at 37°C for 30 min.

    Journal: Frontiers in Chemistry

    Article Title: DNA oxidation profiles of copper phenanthrene chemical nucleases

    doi: 10.3389/fchem.2015.00028

    Figure Lengend Snippet: Lane 1–4 (A–D) DNA cleavage reactions with 250 nM, 500 nM, 1.0 μM, and 2.5 μM test complex (A) Cu-Phen, (B) Cu-DPQ-Phen, (C) Cu-DPPZ-Phen, and (D) Cu-Terph, 400 ng superhelical pUC19 and 1 mM added Na-L-ascorbate incubated at 37°C for 30 min . Lanes 5–16 (A–D) DNA cleavage reactions in the presence of recognition elements, methyl green (MG), netropsin (Net), and [Co(NH 3 ) 6 ]Cl 3 (Co(III)), where 400 ng pUC19 was initially pre-treated with 8 μM of respective non-covalent binding control at 37° C for 45 min and then with 250 nM, 500 nM, 1 μM, and 2.5 μM test complex in the presence of 1 mM added Na-L-ascorbate at 37°C for 30 min.

    Article Snippet: Reactions were carried out according to the following general procedure: in a total volume of 20 μL using 80 mM HEPES buffer (pH 7.2) with 25 mM NaCl, 1 mM Na-L-ascorbate, 400 ng superhelical pUC19 (NEB, N3041) and varying concentrations of test complex (250 nM, 500 nM, 1 μM and 2.5 μM).

    Techniques: Incubation, Binding Assay