puc19  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Name:
    pUC19 Vector
    Description:

    Catalog Number:
    N3041
    Price:
    308
    Category:
    Nucleic Acids
    Applications:
    DNA Manipulation
    Size:
    250 µg
    Buy from Supplier


    Structured Review

    New England Biolabs puc19
    pUC19 Vector

    https://www.bioz.com/result/puc19/product/New England Biolabs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    puc19 - by Bioz Stars, 2021-07
    97/100 stars

    Images

    1) Product Images from "Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974 ▿"

    Article Title: Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974 ▿

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00782-09

    In vivo activities of Streptomyces SerRS1 and SerRS2. s erS1 , s erS2 , E. coli s erS , and the s erS2 ( H270G ) mutant were cloned into pUC19 (pUC) vector under the control of the glnS ). (A) The plasmids were transformed into an E. coli temperature-sensitive
    Figure Legend Snippet: In vivo activities of Streptomyces SerRS1 and SerRS2. s erS1 , s erS2 , E. coli s erS , and the s erS2 ( H270G ) mutant were cloned into pUC19 (pUC) vector under the control of the glnS ). (A) The plasmids were transformed into an E. coli temperature-sensitive

    Techniques Used: In Vivo, Mutagenesis, Clone Assay, Plasmid Preparation, Transformation Assay

    2) Product Images from "C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents"

    Article Title: C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkw1097

    ( A ) Influence of ionic strength on pUC19 condensation (400 ng) by OC3 and MC3 (25 μM) opioid compounds. Condensation reactions on pUC19 (400 ng) by opioid compounds in ( B ) acidic NaOAc buffer (80 mM, pH = 4.0), and ( C ) basic Tris buffer (80 mM, pH = 9.0) in the presence of 25 mM NaCl.
    Figure Legend Snippet: ( A ) Influence of ionic strength on pUC19 condensation (400 ng) by OC3 and MC3 (25 μM) opioid compounds. Condensation reactions on pUC19 (400 ng) by opioid compounds in ( B ) acidic NaOAc buffer (80 mM, pH = 4.0), and ( C ) basic Tris buffer (80 mM, pH = 9.0) in the presence of 25 mM NaCl.

    Techniques Used:

    ( A ) Agarose gel electrophoresis of supercoiled (400 ng) and ( B ) a 742 bp dsDNA fragment of pUC19 (400 ng) exposed to increasing concentrations of MC3, OC3 and HC3 . Reactions were carried out in the presence of 25 mM NaCl for 5 h at 37°C prior to electrophoretic analysis.
    Figure Legend Snippet: ( A ) Agarose gel electrophoresis of supercoiled (400 ng) and ( B ) a 742 bp dsDNA fragment of pUC19 (400 ng) exposed to increasing concentrations of MC3, OC3 and HC3 . Reactions were carried out in the presence of 25 mM NaCl for 5 h at 37°C prior to electrophoretic analysis.

    Techniques Used: Agarose Gel Electrophoresis

    Atomic force microscopy (AFM) images of MC3 -treated supercoiled and HindIII linearized pUC19 DNA; ( A–D ) supercoiled pUC19 with 8, 9, 10 and 20 μM MC3 ; ( E–H ) linear pUC19 with 5, 10, 20 and 50 μM MC3 .
    Figure Legend Snippet: Atomic force microscopy (AFM) images of MC3 -treated supercoiled and HindIII linearized pUC19 DNA; ( A–D ) supercoiled pUC19 with 8, 9, 10 and 20 μM MC3 ; ( E–H ) linear pUC19 with 5, 10, 20 and 50 μM MC3 .

    Techniques Used: Microscopy

    3) Product Images from "Rpn (YhgA-Like) Proteins of Escherichia coli K-12 and Their Contribution to RecA-Independent Horizontal Transfer"

    Article Title: Rpn (YhgA-Like) Proteins of Escherichia coli K-12 and Their Contribution to RecA-Independent Horizontal Transfer

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00787-16

    In vitro analysis of RpnA endonuclease activity. (A) WT RpnA cleaves pUC19, RpnA-D63A does not cleave pUC19, and RpnA-D165A is more active on pUC19. The pUC19 DNA (29 nM, 50 μg/ml) is initially supercoiled but can be relaxed by nicks, linearized by double-strand cleavage, or cleaved further. The supercoiled (control), relaxed (Nb.BtsI), and linear (HindIII) forms are indicated. pUC19 was treated with RpnA-inactive RpnA-D63A or hyperactive RpnA-D165A (15 μM, 45 min). (B) Time course of an RpnA (7.5 μM)-pUC19 (29 nM) digest. Band intensity was compared to determine the relative amounts of supercoiled, nicked, and linear pUC19 at each time point. Over 90% of the supercoiled pUC19 was digested within 180 min. (C) RpnA endonuclease activity depends on divalent cation and is stimulated by Ca 2+ . The reaction buffer was 50 mM NaCl and 10 mM Tris, pH 8.0; the indicated additives were at 10 mM each. RpnA at 3.8 μM was added for 18 h. (D) RpnA cleavage products provide a DNA polymerase primer. pUC19 was digested with RpnA, DNase I, or micrococcal nuclease (MNase) to produce similar smears and then incubated with fluorescein-labeled dNTPs and the Klenow fragment of DNA polymerase. DNA was visualized by ethidium bromide (EtBr; left) or fluorescein (middle), with the two signals being merged at the right. RpnA- and DNase I-digested DNAs were effectively labeled, but micrococcal nuclease-digested DNA was not.
    Figure Legend Snippet: In vitro analysis of RpnA endonuclease activity. (A) WT RpnA cleaves pUC19, RpnA-D63A does not cleave pUC19, and RpnA-D165A is more active on pUC19. The pUC19 DNA (29 nM, 50 μg/ml) is initially supercoiled but can be relaxed by nicks, linearized by double-strand cleavage, or cleaved further. The supercoiled (control), relaxed (Nb.BtsI), and linear (HindIII) forms are indicated. pUC19 was treated with RpnA-inactive RpnA-D63A or hyperactive RpnA-D165A (15 μM, 45 min). (B) Time course of an RpnA (7.5 μM)-pUC19 (29 nM) digest. Band intensity was compared to determine the relative amounts of supercoiled, nicked, and linear pUC19 at each time point. Over 90% of the supercoiled pUC19 was digested within 180 min. (C) RpnA endonuclease activity depends on divalent cation and is stimulated by Ca 2+ . The reaction buffer was 50 mM NaCl and 10 mM Tris, pH 8.0; the indicated additives were at 10 mM each. RpnA at 3.8 μM was added for 18 h. (D) RpnA cleavage products provide a DNA polymerase primer. pUC19 was digested with RpnA, DNase I, or micrococcal nuclease (MNase) to produce similar smears and then incubated with fluorescein-labeled dNTPs and the Klenow fragment of DNA polymerase. DNA was visualized by ethidium bromide (EtBr; left) or fluorescein (middle), with the two signals being merged at the right. RpnA- and DNase I-digested DNAs were effectively labeled, but micrococcal nuclease-digested DNA was not.

    Techniques Used: In Vitro, Activity Assay, Incubation, Labeling

    4) Product Images from "C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents"

    Article Title: C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkw1097

    ( A ) Influence of ionic strength on pUC19 condensation (400 ng) by OC3 and MC3 (25 μM) opioid compounds. Condensation reactions on pUC19 (400 ng) by opioid compounds in ( B ) acidic NaOAc buffer (80 mM, pH = 4.0), and ( C ) basic Tris buffer (80 mM, pH = 9.0) in the presence of 25 mM NaCl.
    Figure Legend Snippet: ( A ) Influence of ionic strength on pUC19 condensation (400 ng) by OC3 and MC3 (25 μM) opioid compounds. Condensation reactions on pUC19 (400 ng) by opioid compounds in ( B ) acidic NaOAc buffer (80 mM, pH = 4.0), and ( C ) basic Tris buffer (80 mM, pH = 9.0) in the presence of 25 mM NaCl.

    Techniques Used:

    ( A ) Agarose gel electrophoresis of supercoiled (400 ng) and ( B ) a 742 bp dsDNA fragment of pUC19 (400 ng) exposed to increasing concentrations of MC3, OC3 and HC3 . Reactions were carried out in the presence of 25 mM NaCl for 5 h at 37°C prior to electrophoretic analysis.
    Figure Legend Snippet: ( A ) Agarose gel electrophoresis of supercoiled (400 ng) and ( B ) a 742 bp dsDNA fragment of pUC19 (400 ng) exposed to increasing concentrations of MC3, OC3 and HC3 . Reactions were carried out in the presence of 25 mM NaCl for 5 h at 37°C prior to electrophoretic analysis.

    Techniques Used: Agarose Gel Electrophoresis

    Atomic force microscopy (AFM) images of MC3 -treated supercoiled and HindIII linearized pUC19 DNA; ( A–D ) supercoiled pUC19 with 8, 9, 10 and 20 μM MC3 ; ( E–H ) linear pUC19 with 5, 10, 20 and 50 μM MC3 .
    Figure Legend Snippet: Atomic force microscopy (AFM) images of MC3 -treated supercoiled and HindIII linearized pUC19 DNA; ( A–D ) supercoiled pUC19 with 8, 9, 10 and 20 μM MC3 ; ( E–H ) linear pUC19 with 5, 10, 20 and 50 μM MC3 .

    Techniques Used: Microscopy

    5) Product Images from "Engineering Nt.BtsCI and Nb.BtsCI nicking enzymes and applications in generating long overhangs"

    Article Title: Engineering Nt.BtsCI and Nb.BtsCI nicking enzymes and applications in generating long overhangs

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkp1092

    DNA nicking activity of top-strand nicking BtsCI variant. Two-fold serial dilutions of the cell extract of BtsCI nicking variant E128F were incubated with pUC19 as described in ‘Materials and Methods’ section. The cleavage products were analyzed on a 1% agarose gel. OC, open circle; SC, supercoiled.
    Figure Legend Snippet: DNA nicking activity of top-strand nicking BtsCI variant. Two-fold serial dilutions of the cell extract of BtsCI nicking variant E128F were incubated with pUC19 as described in ‘Materials and Methods’ section. The cleavage products were analyzed on a 1% agarose gel. OC, open circle; SC, supercoiled.

    Techniques Used: Activity Assay, Variant Assay, Incubation, Agarose Gel Electrophoresis

    DNA nicking activity of the bottom-strand nicking BtsCI variant. Two-fold serial dilutions of the crude extract of BtsCI nicking variant (D388A/E403A/E405A) were incubated with pUC19 as described in ‘Materials and Methods’ section. The cleavage products were analyzed on a 1% agarose gel. OC, open circle; SC, supercoiled.
    Figure Legend Snippet: DNA nicking activity of the bottom-strand nicking BtsCI variant. Two-fold serial dilutions of the crude extract of BtsCI nicking variant (D388A/E403A/E405A) were incubated with pUC19 as described in ‘Materials and Methods’ section. The cleavage products were analyzed on a 1% agarose gel. OC, open circle; SC, supercoiled.

    Techniques Used: Activity Assay, Variant Assay, Incubation, Agarose Gel Electrophoresis

    DNA nicking activity of BtsCI mutants. Two-fold serial dilutions of the clarified cell extracts of E. coli cultures that expressed the indicated BtsCI mutants were incubated with 0.5 µg of pUC19 as described in ‘Materials and Methods’ section. The cleavage products were analyzed on a 1% agarose gel. OC, open circle; SC, supercoiled; −, no cleavage; +, pUC19 nicked by Nt.BsmAI.
    Figure Legend Snippet: DNA nicking activity of BtsCI mutants. Two-fold serial dilutions of the clarified cell extracts of E. coli cultures that expressed the indicated BtsCI mutants were incubated with 0.5 µg of pUC19 as described in ‘Materials and Methods’ section. The cleavage products were analyzed on a 1% agarose gel. OC, open circle; SC, supercoiled; −, no cleavage; +, pUC19 nicked by Nt.BsmAI.

    Techniques Used: Activity Assay, Incubation, Agarose Gel Electrophoresis

    ( A ) Generation of overhangs. Cleavage sites for BtsCI, FokI and a combination of FokI/Nt.BtsCI and FokI/Nb.BtsCI are indicated. For BtsCI, a 2-nt 5′ recessive end is generated. For FokI, a 4-nt 3′ recessive end is generated. For FokI/Nt.BtsCI, an 11-nt 3′ recessive end is generated. For FokI/Nb.BtsCI, a 9-nt 5′ recessive end is generated. Grey arrows, BtsCI top-strand cleavage site; white arrows, BtsCI bottom-strand cleavage site; black arrows, FokI cleavage sites. ( B ) Annealing of oligonucleotides to the long overhangs for PCR. Plasmid pUC19 was cleaved by the indicated enzyme(s) and then ligated to a 100-bp long oligonucleotide as described in ‘Materials and Methods’ section. The ligation products were used as template for PCR that specifically detect the ligated DNA. Only the cleavage product of FokI/Nt.BtsCI can be annealed to the 11-nt 3′ recessive end oligonucleotide (left panel), whereas only the cleavage product of FokI/Nb.BtsCI can be annealed to the 9-nt 5′ recessive end oligonucleotide (right panel).
    Figure Legend Snippet: ( A ) Generation of overhangs. Cleavage sites for BtsCI, FokI and a combination of FokI/Nt.BtsCI and FokI/Nb.BtsCI are indicated. For BtsCI, a 2-nt 5′ recessive end is generated. For FokI, a 4-nt 3′ recessive end is generated. For FokI/Nt.BtsCI, an 11-nt 3′ recessive end is generated. For FokI/Nb.BtsCI, a 9-nt 5′ recessive end is generated. Grey arrows, BtsCI top-strand cleavage site; white arrows, BtsCI bottom-strand cleavage site; black arrows, FokI cleavage sites. ( B ) Annealing of oligonucleotides to the long overhangs for PCR. Plasmid pUC19 was cleaved by the indicated enzyme(s) and then ligated to a 100-bp long oligonucleotide as described in ‘Materials and Methods’ section. The ligation products were used as template for PCR that specifically detect the ligated DNA. Only the cleavage product of FokI/Nt.BtsCI can be annealed to the 11-nt 3′ recessive end oligonucleotide (left panel), whereas only the cleavage product of FokI/Nb.BtsCI can be annealed to the 9-nt 5′ recessive end oligonucleotide (right panel).

    Techniques Used: Generated, Polymerase Chain Reaction, Plasmid Preparation, Ligation

    Related Articles

    Purification:

    Article Title: Methods and Techniques to Facilitate the Development of Clostridium novyi NT as an Effective, Therapeutic Oncolytic Bacteria
    Article Snippet: Transformation of Calcium Competent C. novyi Calcium competent C. novyi cells were allowed to thaw on ice. .. Subsequently, 5 μg of either purified pUC19 control plasmid (New England BioLabs Inc.) or the purified pKMD002 plasmid was added to an empty prechilled 15 mL tube (4°C). .. Competent C. novyi cells (100 μL) were added directly on top of the plasmid DNA in the prechilled tube without vortexing or mixing.

    Plasmid Preparation:

    Article Title: Methods and Techniques to Facilitate the Development of Clostridium novyi NT as an Effective, Therapeutic Oncolytic Bacteria
    Article Snippet: Transformation of Calcium Competent C. novyi Calcium competent C. novyi cells were allowed to thaw on ice. .. Subsequently, 5 μg of either purified pUC19 control plasmid (New England BioLabs Inc.) or the purified pKMD002 plasmid was added to an empty prechilled 15 mL tube (4°C). .. Competent C. novyi cells (100 μL) were added directly on top of the plasmid DNA in the prechilled tube without vortexing or mixing.

    Article Title: Base-resolution detection of N4-methylcytosine in genomic DNA using 4mC-Tet-assisted-bisulfite- sequencing
    Article Snippet: .. The 4mC control is generated by PCR amplification with 4mdCTP using part of the pUC19 vector (∼1 kb) as a template, whereas the C/5mC control is generated by treating unmethylated lambda DNA with the CpG methyltransferase (M. SssI ) (Figure ). .. The lambda DNA therefore contains methylated 5mC in the CpG context and unmethylated non-CpG, which is used to measure the efficiency of the bisulfite-mediated conversion and Tet oxidation.

    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: CRISPR-Cas9-Guided Genome Engineering in C. elegans
    Article Snippet: Cas9 Expression Plasmid (Addgene plasmid #46168). pHKMC1 - Empty sgRNA Cloning Plasmid (Addgene plasmid #67720). pCFJ90 - P myo-2::mCherry::unc-54utr (Addgene plasmid #19327). pCFJ104 - P myo-3::mCherry::unc-54 (Addgene plasmid #19328). pMA122 - peel-1 negative selection (Addgene plasmid #34873, Optional). .. P eft-3 :: Cas9-SV40_NLS :: tbb-2 3’UTR (Addgene plasmid #46168). pUC19 (NEB N3041S). pPV477 (Addgene plasmid #42930). .. P myo-2::GFP ).

    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: Reconstruction of cysteine biosynthesis using engineered cysteine-free enzymes
    Article Snippet: .. Two control experiments were also conducted: a positive control consisting of electrocompetent ΔcysE cells transformed with cysE in pUC19 vector and the other with electrocompetent ΔcysKΔcysM cells transformed with codon optimized cysM in pUC19 vector. .. The second positive control group supplemented the lack of cysteine through growing the knockout cells with empty pUC19 vectors on a fully supplemented media, LB + Amp, and were allowed to incubate at 37 °C overnight.

    Article Title: Ultra-Low Background DNA Cloning System
    Article Snippet: Custom PCR primers (pGFPuv1 and pGFPuv2) purified by polyacrylamide gel electrophoresis were purchased from Hokkaido System Science (Sapporo, Hokkaido, Japan). .. Yeast and E. coli transformation For cloning experiments, the pUC19 vector was digested with Eco RI-HF (New England Biolabs, Ipswich, MI, USA) and Xba I (Fermentas, Vilnius, Lithuania) in the multiple cloning site (MCS). .. The digested vector was purified by MonoFas DNA Purification Kit I.

    Generated:

    Article Title: Base-resolution detection of N4-methylcytosine in genomic DNA using 4mC-Tet-assisted-bisulfite- sequencing
    Article Snippet: .. The 4mC control is generated by PCR amplification with 4mdCTP using part of the pUC19 vector (∼1 kb) as a template, whereas the C/5mC control is generated by treating unmethylated lambda DNA with the CpG methyltransferase (M. SssI ) (Figure ). .. The lambda DNA therefore contains methylated 5mC in the CpG context and unmethylated non-CpG, which is used to measure the efficiency of the bisulfite-mediated conversion and Tet oxidation.

    Polymerase Chain Reaction:

    Article Title: Base-resolution detection of N4-methylcytosine in genomic DNA using 4mC-Tet-assisted-bisulfite- sequencing
    Article Snippet: .. The 4mC control is generated by PCR amplification with 4mdCTP using part of the pUC19 vector (∼1 kb) as a template, whereas the C/5mC control is generated by treating unmethylated lambda DNA with the CpG methyltransferase (M. SssI ) (Figure ). .. The lambda DNA therefore contains methylated 5mC in the CpG context and unmethylated non-CpG, which is used to measure the efficiency of the bisulfite-mediated conversion and Tet oxidation.

    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: .. The 4mC control is generated by PCR amplification with 4mdCTP using part of the pUC19 vector (∼1 kb) as a template, whereas the C/5mC control is generated by treating unmethylated lambda DNA with the CpG methyltransferase (M. SssI ) (Figure ). .. The lambda DNA therefore contains methylated 5mC in the CpG context and unmethylated non-CpG, which is used to measure the efficiency of the bisulfite-mediated conversion and Tet oxidation.

    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). ..

    Lambda DNA Preparation:

    Article Title: Base-resolution detection of N4-methylcytosine in genomic DNA using 4mC-Tet-assisted-bisulfite- sequencing
    Article Snippet: .. The 4mC control is generated by PCR amplification with 4mdCTP using part of the pUC19 vector (∼1 kb) as a template, whereas the C/5mC control is generated by treating unmethylated lambda DNA with the CpG methyltransferase (M. SssI ) (Figure ). .. The lambda DNA therefore contains methylated 5mC in the CpG context and unmethylated non-CpG, which is used to measure the efficiency of the bisulfite-mediated conversion and Tet oxidation.

    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: Ultra-Low Background DNA Cloning System
    Article Snippet: Custom PCR primers (pGFPuv1 and pGFPuv2) purified by polyacrylamide gel electrophoresis were purchased from Hokkaido System Science (Sapporo, Hokkaido, Japan). .. Yeast and E. coli transformation For cloning experiments, the pUC19 vector was digested with Eco RI-HF (New England Biolabs, Ipswich, MI, USA) and Xba I (Fermentas, Vilnius, Lithuania) in the multiple cloning site (MCS). .. The digested vector was purified by MonoFas DNA Purification Kit I.

    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: Reconstruction of cysteine biosynthesis using engineered cysteine-free enzymes
    Article Snippet: .. Two control experiments were also conducted: a positive control consisting of electrocompetent ΔcysE cells transformed with cysE in pUC19 vector and the other with electrocompetent ΔcysKΔcysM cells transformed with codon optimized cysM in pUC19 vector. .. The second positive control group supplemented the lack of cysteine through growing the knockout cells with empty pUC19 vectors on a fully supplemented media, LB + Amp, and were allowed to incubate at 37 °C overnight.

    Article Title: Ultra-Low Background DNA Cloning System
    Article Snippet: Custom PCR primers (pGFPuv1 and pGFPuv2) purified by polyacrylamide gel electrophoresis were purchased from Hokkaido System Science (Sapporo, Hokkaido, Japan). .. Yeast and E. coli transformation For cloning experiments, the pUC19 vector was digested with Eco RI-HF (New England Biolabs, Ipswich, MI, USA) and Xba I (Fermentas, Vilnius, Lithuania) in the multiple cloning site (MCS). .. The digested vector was purified by MonoFas DNA Purification Kit I.

    Incubation:

    Article Title: C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents
    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, 400 ng pUC19 (NEB, N3041) and varying concentrations of test compound (5, 10, 20 and 30 μM), samples were incubated at 37°C for both 5 and 12 h. Reactions were quenched by adding 6x loading buffer (Fermentas) containing 10 mM Tris-HCl, 0.03% bromophenol blue, 0.03% xylene cyanole FF, 60% glycerol, 60 mM EDTA and samples were loaded onto an agarose gel (1.2%) containing 3 μl EtBr. .. Electrophoresis was completed at 60 V for 1 h in 1x TAE buffer.

    Agarose Gel Electrophoresis:

    Article Title: C3-symmetric opioid scaffolds are pH-responsive DNA condensation agents
    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, 400 ng pUC19 (NEB, N3041) and varying concentrations of test compound (5, 10, 20 and 30 μM), samples were incubated at 37°C for both 5 and 12 h. Reactions were quenched by adding 6x loading buffer (Fermentas) containing 10 mM Tris-HCl, 0.03% bromophenol blue, 0.03% xylene cyanole FF, 60% glycerol, 60 mM EDTA and samples were loaded onto an agarose gel (1.2%) containing 3 μl EtBr. .. Electrophoresis was completed at 60 V for 1 h in 1x TAE buffer.

    Positive Control:

    Article Title: Reconstruction of cysteine biosynthesis using engineered cysteine-free enzymes
    Article Snippet: .. Two control experiments were also conducted: a positive control consisting of electrocompetent ΔcysE cells transformed with cysE in pUC19 vector and the other with electrocompetent ΔcysKΔcysM cells transformed with codon optimized cysM in pUC19 vector. .. The second positive control group supplemented the lack of cysteine through growing the knockout cells with empty pUC19 vectors on a fully supplemented media, LB + Amp, and were allowed to incubate at 37 °C overnight.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    New England Biolabs puc19 vector dna
    Data analysis of spike-in controls from MethylC-seq and 4mC-TAB-seq in the context of C. kristjanssonii genomic <t>DNA.</t> ( A ) Composition of <t>pUC19</t> DNA, lambda DNA, and C. kristjanssonii genomic DNA. ( B ) The percentage of detected as cytosine reads on 4mC sites in untreated and Tet-treated samples. ( C ) The detected as cytosine reads percentage on unmodified cytosine sites (non-CpG context) and 5mC sites (CpG context) in untreated and Tet-treated samples. ( D ) Quantification of 4mC and 5mC in C. kristjanssonii genomic DNA, determined by LC-MS/MS and deep-sequencing respectively. Error bars indicate mean ± SD, n = 4.
    Puc19 Vector Dna, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/puc19 vector dna/product/New England Biolabs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    puc19 vector dna - by Bioz Stars, 2021-07
    97/100 stars
      Buy from Supplier

    99
    New England Biolabs puc19 vector
    Synthetic c ysE and cysM gene transformants display recovery of CysE function without cysteine and methionine and CysM function without cysteine. ( A ) E. coli ΔcysE competent cells were transformed with positive control cysE , two cysE variants cysE-C/cysE-CM cloned into the multiple cloning site of <t>pUC19</t> plasmid, and original pUC19 encoding N-terminal fragment of lacZ α as a negative control. ( B ) E. coli ΔcysMΔcysK competent cells transformed with positive control c ysM , two cysM variants cysM-C / cysM-CM in pUC19 plasmid, and original pUC19 encoding N-terminal fragment of lacZ α as a negative control. Cells were plated on M9 + glucose medium with 0.4 mM IPTG, 50 μg/ml kanamycin, and 100 μg/ml ampicillin and incubated at 30 °C for 72 h.
    Puc19 Vector, 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/puc19 vector/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    puc19 vector - by Bioz Stars, 2021-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    Data analysis of spike-in controls from MethylC-seq and 4mC-TAB-seq in the context of C. kristjanssonii genomic DNA. ( A ) Composition of pUC19 DNA, lambda DNA, and C. kristjanssonii genomic DNA. ( B ) The percentage of detected as cytosine reads on 4mC sites in untreated and Tet-treated samples. ( C ) The detected as cytosine reads percentage on unmodified cytosine sites (non-CpG context) and 5mC sites (CpG context) in untreated and Tet-treated samples. ( D ) Quantification of 4mC and 5mC in C. kristjanssonii genomic DNA, determined by LC-MS/MS and deep-sequencing respectively. Error bars indicate mean ± SD, n = 4.

    Journal: Nucleic Acids Research

    Article Title: Base-resolution detection of N4-methylcytosine in genomic DNA using 4mC-Tet-assisted-bisulfite- sequencing

    doi: 10.1093/nar/gkv738

    Figure Lengend Snippet: Data analysis of spike-in controls from MethylC-seq and 4mC-TAB-seq in the context of C. kristjanssonii genomic DNA. ( A ) Composition of pUC19 DNA, lambda DNA, and C. kristjanssonii genomic DNA. ( B ) The percentage of detected as cytosine reads on 4mC sites in untreated and Tet-treated samples. ( C ) The detected as cytosine reads percentage on unmodified cytosine sites (non-CpG context) and 5mC sites (CpG context) in untreated and Tet-treated samples. ( D ) Quantification of 4mC and 5mC in C. kristjanssonii genomic DNA, determined by LC-MS/MS and deep-sequencing respectively. Error bars indicate mean ± SD, n = 4.

    Article Snippet: 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).

    Techniques: Lambda DNA Preparation, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Sequencing

    Calcium Competent C. novyi Transformations. (A) Schematic representation of experimental flow. (B) Resulting colony forming units after Clostridium novyi cells underwent calcium competent transformation with the E. coli plasmid pUC19, which contains a gene encoding ampicillin resistance. Statistical significance was determined to be p

    Journal: Frontiers in Microbiology

    Article Title: Methods and Techniques to Facilitate the Development of Clostridium novyi NT as an Effective, Therapeutic Oncolytic Bacteria

    doi: 10.3389/fmicb.2021.624618

    Figure Lengend Snippet: Calcium Competent C. novyi Transformations. (A) Schematic representation of experimental flow. (B) Resulting colony forming units after Clostridium novyi cells underwent calcium competent transformation with the E. coli plasmid pUC19, which contains a gene encoding ampicillin resistance. Statistical significance was determined to be p

    Article Snippet: Subsequently, 5 μg of either purified pUC19 control plasmid (New England BioLabs Inc.) or the purified pKMD002 plasmid was added to an empty prechilled 15 mL tube (4°C).

    Techniques: Transformation Assay, Plasmid Preparation

    In vivo activities of Streptomyces SerRS1 and SerRS2. s erS1 , s erS2 , E. coli s erS , and the s erS2 ( H270G ) mutant were cloned into pUC19 (pUC) vector under the control of the glnS ). (A) The plasmids were transformed into an E. coli temperature-sensitive

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974 ▿

    doi: 10.1128/AAC.00782-09

    Figure Lengend Snippet: In vivo activities of Streptomyces SerRS1 and SerRS2. s erS1 , s erS2 , E. coli s erS , and the s erS2 ( H270G ) mutant were cloned into pUC19 (pUC) vector under the control of the glnS ). (A) The plasmids were transformed into an E. coli temperature-sensitive

    Article Snippet: 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.

    Techniques: In Vivo, Mutagenesis, Clone Assay, Plasmid Preparation, Transformation Assay

    Synthetic c ysE and cysM gene transformants display recovery of CysE function without cysteine and methionine and CysM function without cysteine. ( A ) E. coli ΔcysE competent cells were transformed with positive control cysE , two cysE variants cysE-C/cysE-CM cloned into the multiple cloning site of pUC19 plasmid, and original pUC19 encoding N-terminal fragment of lacZ α as a negative control. ( B ) E. coli ΔcysMΔcysK competent cells transformed with positive control c ysM , two cysM variants cysM-C / cysM-CM in pUC19 plasmid, and original pUC19 encoding N-terminal fragment of lacZ α as a negative control. Cells were plated on M9 + glucose medium with 0.4 mM IPTG, 50 μg/ml kanamycin, and 100 μg/ml ampicillin and incubated at 30 °C for 72 h.

    Journal: Scientific Reports

    Article Title: Reconstruction of cysteine biosynthesis using engineered cysteine-free enzymes

    doi: 10.1038/s41598-018-19920-y

    Figure Lengend Snippet: Synthetic c ysE and cysM gene transformants display recovery of CysE function without cysteine and methionine and CysM function without cysteine. ( A ) E. coli ΔcysE competent cells were transformed with positive control cysE , two cysE variants cysE-C/cysE-CM cloned into the multiple cloning site of pUC19 plasmid, and original pUC19 encoding N-terminal fragment of lacZ α as a negative control. ( B ) E. coli ΔcysMΔcysK competent cells transformed with positive control c ysM , two cysM variants cysM-C / cysM-CM in pUC19 plasmid, and original pUC19 encoding N-terminal fragment of lacZ α as a negative control. Cells were plated on M9 + glucose medium with 0.4 mM IPTG, 50 μg/ml kanamycin, and 100 μg/ml ampicillin and incubated at 30 °C for 72 h.

    Article Snippet: For cloning and transformation, both the PCR products and the pUC19 vector (New England Biolabs) were digested using HindIII-HF and XhoI restriction enzymes at 37 °C for 1 h in 1x CutSmart buffer (New England Biolabs).

    Techniques: Transformation Assay, Positive Control, Clone Assay, Plasmid Preparation, Negative Control, Incubation