ncoi restriction enzymes  (New England Biolabs)


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    Structured Review

    New England Biolabs ncoi restriction enzymes
    pET28a-baPrs-hdNadV bicistronic vector construction. Prs gene from Bacillus amyloliquefaciens with L135I mutation (ctc to ata) was cloned in pUC57-kan vector (GenScript synthesis). pET28a-hdNadV Vector and PCR amplification of prs fragment were digested (with <t>NcoI</t> and <t>XbaI</t> restriction enzymes) and the fragments corresponding to 1030 bp and 6687 bp were purified and ligated. The DNA construct was chemically transformed in Escherichia coli DH5α, verified by agarose gel electrophoresis and transformed in strain BL21(DE3)pLysS. After transformation, cells were grown into a 500 mL bench-top bioreactor system and the NMN yield was determined by derivatization followed by fluorimetric assay.
    Ncoi Restriction Enzymes, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ncoi restriction enzymes/product/New England Biolabs
    Average 95 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    ncoi restriction enzymes - by Bioz Stars, 2022-11
    95/100 stars

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    1) Product Images from "β-nicotinamide mononucleotide (NMN) production in Escherichia coli"

    Article Title: β-nicotinamide mononucleotide (NMN) production in Escherichia coli

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-30792-0

    pET28a-baPrs-hdNadV bicistronic vector construction. Prs gene from Bacillus amyloliquefaciens with L135I mutation (ctc to ata) was cloned in pUC57-kan vector (GenScript synthesis). pET28a-hdNadV Vector and PCR amplification of prs fragment were digested (with NcoI and XbaI restriction enzymes) and the fragments corresponding to 1030 bp and 6687 bp were purified and ligated. The DNA construct was chemically transformed in Escherichia coli DH5α, verified by agarose gel electrophoresis and transformed in strain BL21(DE3)pLysS. After transformation, cells were grown into a 500 mL bench-top bioreactor system and the NMN yield was determined by derivatization followed by fluorimetric assay.
    Figure Legend Snippet: pET28a-baPrs-hdNadV bicistronic vector construction. Prs gene from Bacillus amyloliquefaciens with L135I mutation (ctc to ata) was cloned in pUC57-kan vector (GenScript synthesis). pET28a-hdNadV Vector and PCR amplification of prs fragment were digested (with NcoI and XbaI restriction enzymes) and the fragments corresponding to 1030 bp and 6687 bp were purified and ligated. The DNA construct was chemically transformed in Escherichia coli DH5α, verified by agarose gel electrophoresis and transformed in strain BL21(DE3)pLysS. After transformation, cells were grown into a 500 mL bench-top bioreactor system and the NMN yield was determined by derivatization followed by fluorimetric assay.

    Techniques Used: Plasmid Preparation, Mutagenesis, Clone Assay, Polymerase Chain Reaction, Amplification, Purification, Construct, Transformation Assay, Agarose Gel Electrophoresis, Fluorimetry Assay

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    New England Biolabs restriction enzyme digestion
    One percent agarose gel. ( A ) Electrophoresis of the PCR product of the Zika virus recombinant protein E (E-ZIKVre) gene in the commercial vector pUCIDT + E-ZIKVre. Lane 1—Molecular marker 1kb DNA Ladder™ (Life Technologies); line 2–4—PCR product E-ZIKVb; lane 5—negative control. ( B ) Electrophoresis of the enzymatic cleavage digestion products of <t>the</t> <t>pET28a</t> + E-ZIKVre construct with <t>restriction</t> <t>endonucleases</t> <t>NcoI</t> and HindIII. Lane 1—Marker 1kb DNA Ladder™ (Life Technologies); lane 2—digested clone 1; lane 3—undigested clone 1; lane 4—digested clone 2; lane 5—undigested clone 2; lane 6—digested clone 3; lane 7—undigested clone 3; lane 8—digested clone 4; lane 9—undigested clone 4; lane 10—digested clone 5; lane 11—undigested clone 5; lane 12—digested clone 6; and lane 13—undigested clone 6.
    Restriction Enzyme Digestion, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    One percent agarose gel. ( A ) Electrophoresis of the PCR product of the Zika virus recombinant protein E (E-ZIKVre) gene in the commercial vector pUCIDT + E-ZIKVre. Lane 1—Molecular marker 1kb DNA Ladder™ (Life Technologies); line 2–4—PCR product E-ZIKVb; lane 5—negative control. ( B ) Electrophoresis of the enzymatic cleavage digestion products of the pET28a + E-ZIKVre construct with restriction endonucleases NcoI and HindIII. Lane 1—Marker 1kb DNA Ladder™ (Life Technologies); lane 2—digested clone 1; lane 3—undigested clone 1; lane 4—digested clone 2; lane 5—undigested clone 2; lane 6—digested clone 3; lane 7—undigested clone 3; lane 8—digested clone 4; lane 9—undigested clone 4; lane 10—digested clone 5; lane 11—undigested clone 5; lane 12—digested clone 6; and lane 13—undigested clone 6.

    Journal: Viruses

    Article Title: Selection and Characterization of Single-Stranded DNA Aptamers of Diagnostic Potential against the Whole Zika Virus

    doi: 10.3390/v14091867

    Figure Lengend Snippet: One percent agarose gel. ( A ) Electrophoresis of the PCR product of the Zika virus recombinant protein E (E-ZIKVre) gene in the commercial vector pUCIDT + E-ZIKVre. Lane 1—Molecular marker 1kb DNA Ladder™ (Life Technologies); line 2–4—PCR product E-ZIKVb; lane 5—negative control. ( B ) Electrophoresis of the enzymatic cleavage digestion products of the pET28a + E-ZIKVre construct with restriction endonucleases NcoI and HindIII. Lane 1—Marker 1kb DNA Ladder™ (Life Technologies); lane 2—digested clone 1; lane 3—undigested clone 1; lane 4—digested clone 2; lane 5—undigested clone 2; lane 6—digested clone 3; lane 7—undigested clone 3; lane 8—digested clone 4; lane 9—undigested clone 4; lane 10—digested clone 5; lane 11—undigested clone 5; lane 12—digested clone 6; and lane 13—undigested clone 6.

    Article Snippet: The E-ZIKVre gene contained in the pUCIDT commercial vector was amplified by PCR with specific oligonucleotides: E-ZIKVre FW (5′-CAT GCC ATG GGC ATT AGG TGC ATA GGC GTT AGC-3′) and E-ZIKVre RV (5′-CCC AAG CTT CTA ATG GTG GTG ATG GTG ATG C-3′) and then cloned into the pET28a expression vector (Novagen, Madison, WI, USA) using restriction endonucleases NcoI (New England Biolabs, Ipswich, MA, USA) and HindIII (New England Biolabs, Ipswich, MA, USA) and the enzyme T4 DNA ligase (Life Technologies, Carlsbad, CA, USA), following the manufacturer’s recommendations.

    Techniques: Agarose Gel Electrophoresis, Electrophoresis, Polymerase Chain Reaction, Recombinant, Plasmid Preparation, Marker, Negative Control, Construct