restriction enzymes ncoi  (New England Biolabs)


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    Structured Review

    New England Biolabs restriction enzymes ncoi
    The Double Digestion Patterns of the pNZ8148-HPV16-optiE7 Shuttle Plasmid via <t>NcoI</t> and <t>SacI</t> Restriction Endonuclease
    Restriction Enzymes Ncoi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction enzymes ncoi/product/New England Biolabs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    restriction enzymes ncoi - by Bioz Stars, 2022-07
    96/100 stars

    Images

    1) Product Images from "Codon Usage Optimization and Construction of Plasmid Encoding Iranian Human Papillomavirus Type 16 E7 Oncogene for Lactococcus Lactis Subsp. Cremoris MG1363"

    Article Title: Codon Usage Optimization and Construction of Plasmid Encoding Iranian Human Papillomavirus Type 16 E7 Oncogene for Lactococcus Lactis Subsp. Cremoris MG1363

    Journal: Asian Pacific Journal of Cancer Prevention : APJCP

    doi: 10.22034/APJCP.2017.18.3.783

    The Double Digestion Patterns of the pNZ8148-HPV16-optiE7 Shuttle Plasmid via NcoI and SacI Restriction Endonuclease
    Figure Legend Snippet: The Double Digestion Patterns of the pNZ8148-HPV16-optiE7 Shuttle Plasmid via NcoI and SacI Restriction Endonuclease

    Techniques Used: Plasmid Preparation

    2) Product Images from "Protein switches identified from diverse insertion libraries created using S1 nuclease digestion of supercoiled-form plasmid DNA"

    Article Title: Protein switches identified from diverse insertion libraries created using S1 nuclease digestion of supercoiled-form plasmid DNA

    Journal: Biotechnology and bioengineering

    doi: 10.1002/bit.23224

    S1 nuclease converts supercoiled plasmid DNA to nicked circular and linear forms through digestion at a variety of locations in the plasmid. (A) Two μg of supercoiled pRH04.152-rbsB was incubated at 22°C (top) or 37°C (bottom) with different amounts of S1 nuclease for different lengths of time and then analyzed by agarose gel electrophoresis. The sixth and seventh lanes contain undigested supercoiled control and SpeI-digested linear control, respectively. The band that runs slower than the linear band was presumed to correspond to nicked circular plasmid. In the thirteenth lane, after the DNA was digested with 10 units for 24 hours an additional 10 units of S1 nuclease was added and the DNA was incubated for an additional hour. The first and last lanes contain the λDNA/HindIII molecular weight standards. (B) Plasmids with or without the f1 origin (pDIMC8-pfMBP and pRH04-pfMBP, respectively) were incubated with or without S1 nuclease, followed by incubation with or without NcoI, EagI, and SacII and then analyzed by agarose gel electrophoresis. (C) Plasmids with or without the f1 origin (pDIMC8-rbsb and pRH04-rbsb, respectively) were incubated with S1 nuclease or DNaseI, followed by incubation with XmnI and analyzed by agarose gel electrophoresis.
    Figure Legend Snippet: S1 nuclease converts supercoiled plasmid DNA to nicked circular and linear forms through digestion at a variety of locations in the plasmid. (A) Two μg of supercoiled pRH04.152-rbsB was incubated at 22°C (top) or 37°C (bottom) with different amounts of S1 nuclease for different lengths of time and then analyzed by agarose gel electrophoresis. The sixth and seventh lanes contain undigested supercoiled control and SpeI-digested linear control, respectively. The band that runs slower than the linear band was presumed to correspond to nicked circular plasmid. In the thirteenth lane, after the DNA was digested with 10 units for 24 hours an additional 10 units of S1 nuclease was added and the DNA was incubated for an additional hour. The first and last lanes contain the λDNA/HindIII molecular weight standards. (B) Plasmids with or without the f1 origin (pDIMC8-pfMBP and pRH04-pfMBP, respectively) were incubated with or without S1 nuclease, followed by incubation with or without NcoI, EagI, and SacII and then analyzed by agarose gel electrophoresis. (C) Plasmids with or without the f1 origin (pDIMC8-rbsb and pRH04-rbsb, respectively) were incubated with S1 nuclease or DNaseI, followed by incubation with XmnI and analyzed by agarose gel electrophoresis.

    Techniques Used: Plasmid Preparation, Incubation, Agarose Gel Electrophoresis, Molecular Weight

    3) Product Images from "The PRESAT-vector: Asymmetric T-vector for high-throughput screening of soluble protein domains for structural proteomics"

    Article Title: The PRESAT-vector: Asymmetric T-vector for high-throughput screening of soluble protein domains for structural proteomics

    Journal: Protein Science : A Publication of the Protein Society

    doi: 10.1110/ps.03439004

    Concept of the asymmetric directional T-vector. ( A ) Construction of pGEX-4T3-PRESAT and direct cloning of PCR product in pGEX-4T3-PRESAT. ( B ) The schematic representation of the ORF selection method using potential restriction enzyme site. The figure illustrates the case in which NcoI is chosen as the second restriction enzyme for selection. The rear PCR primer is designed with 5′-GG at the 5′ end, so that only the ligated plasmid with insert in the reverse orientation will have the NcoI site at the TA-cloning position. For NdeI selection, the rear primer with 5′-ATG is used instead of the 5′-GG primer.
    Figure Legend Snippet: Concept of the asymmetric directional T-vector. ( A ) Construction of pGEX-4T3-PRESAT and direct cloning of PCR product in pGEX-4T3-PRESAT. ( B ) The schematic representation of the ORF selection method using potential restriction enzyme site. The figure illustrates the case in which NcoI is chosen as the second restriction enzyme for selection. The rear PCR primer is designed with 5′-GG at the 5′ end, so that only the ligated plasmid with insert in the reverse orientation will have the NcoI site at the TA-cloning position. For NdeI selection, the rear primer with 5′-ATG is used instead of the 5′-GG primer.

    Techniques Used: Plasmid Preparation, Clone Assay, Polymerase Chain Reaction, Selection, TA Cloning

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    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 1 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 1 article reviews
    Price from $9.99 to $1999.99
    ncoi restriction enzymes - by Bioz Stars, 2022-07
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    96
    New England Biolabs restriction enzymes ncoi
    The Double Digestion Patterns of the pNZ8148-HPV16-optiE7 Shuttle Plasmid via <t>NcoI</t> and <t>SacI</t> Restriction Endonuclease
    Restriction Enzymes Ncoi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction enzymes ncoi/product/New England Biolabs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    restriction enzymes ncoi - by Bioz Stars, 2022-07
    96/100 stars
      Buy from Supplier

    Image Search Results


    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.

    Journal: Scientific Reports

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

    doi: 10.1038/s41598-018-30792-0

    Figure Lengend 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.

    Article Snippet: For the simultaneous expression of Nampt and PRPP synthetase a bicistronic vector was constructed by PCR amplifying the baPrs sequence (Q5 High-Fidelity 2X Master Mix, NEB) from pUC57-Kan plasmid with M13 forward and reverse primers, double digestion with XbaI and NcoI restriction enzymes (NEB) of PCR product and pET28a-hdNAdV plasmid, followed by the separation of the desired DNA fragments on 1.5% agarose gel electrophoresis , purification from gel (using Wizard SV Gel and PCR Clean-Up System, Promega, Madison, USA) of desired bands (pET28a-hdNadV and baPrs) followed by ligation with T4 DNA ligase (NEB) (as shown in Fig. ).

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

    The Double Digestion Patterns of the pNZ8148-HPV16-optiE7 Shuttle Plasmid via NcoI and SacI Restriction Endonuclease

    Journal: Asian Pacific Journal of Cancer Prevention : APJCP

    Article Title: Codon Usage Optimization and Construction of Plasmid Encoding Iranian Human Papillomavirus Type 16 E7 Oncogene for Lactococcus Lactis Subsp. Cremoris MG1363

    doi: 10.22034/APJCP.2017.18.3.783

    Figure Lengend Snippet: The Double Digestion Patterns of the pNZ8148-HPV16-optiE7 Shuttle Plasmid via NcoI and SacI Restriction Endonuclease

    Article Snippet: Construction of shuttle vector The optimized E7 gene, encoding the E7 oncoprotein from HPV 16, was obtained as a 291 bp DNA fragment by digesting plasmid PMD18 with restriction enzymes NcoI and SacI (New England Biolabs).

    Techniques: Plasmid Preparation

    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.

    Journal: Scientific Reports

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

    doi: 10.1038/s41598-018-30792-0

    Figure Lengend 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.

    Article Snippet: For the simultaneous expression of Nampt and PRPP synthetase a bicistronic vector was constructed by PCR amplifying the baPrs sequence (Q5 High-Fidelity 2X Master Mix, NEB) from pUC57-Kan plasmid with M13 forward and reverse primers, double digestion with XbaI and NcoI restriction enzymes (NEB) of PCR product and pET28a-hdNAdV plasmid, followed by the separation of the desired DNA fragments on 1.5% agarose gel electrophoresis , purification from gel (using Wizard SV Gel and PCR Clean-Up System, Promega, Madison, USA) of desired bands (pET28a-hdNadV and baPrs) followed by ligation with T4 DNA ligase (NEB) (as shown in Fig. ).

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

    An interaction between the distal and proximal regulatory regions of the MCP - 1 gene forms following TNF stimulation. (A) A schematic representation of the 3C assay and the architecture of the MCP - 1 promoter with cis regulatory sites with selected restriction sites and primer positions (P1 through P4) is shown. In the 3C assay, cells were fixed with formaldehyde and chromatin was isolated and digested with NcoI. Following inactivation of the restriction enzyme, the sample was diluted and subjected to DNA ligation such that only intramolecular ligations would be preferred. The cross-links were then reversed, and the DNA was purified. PCR primers P1 and P2 were used to detect the formation of the novel ligated 3C product. Primers P3 and P4 were used to verify that similar levels of MCP - 1 DNA were present in the assay and that the DNA was amplifiable. Products from the 3C assay were examined on agarose gels stained with ethidium bromide. (B) NIH 3T3 cells treated in the absence (−) (lanes 1 to 4) or presence (+) (lanes 5 to 8) of TNF for 2 h were processed in the above 3C assay with and without formaldehyde (CH 2 O) or DNA ligase, as indicated, to control for specificity of the protocol. M, DNA marker. (C) Purified mouse genomic DNA was processed in the 3C assay and demonstrates that the unique PCR product cannot form with naked DNA templates. (D) KpnI cleaves the PCR product generated by the 3C assay, producing the anticipated DNA fragments as shown on the agarose gel. (E) PCR amplification of the 3C product and the non-3C product display similar relative efficiencies. PCR products generated from undigested genomic DNA (1,207 bp) and DNA from a 3C assay (713 bp) were quantitated by fluorometry and used as templates in PCRs to determine if there were major differences in their PCR efficiency. DNA templates (40 pg, 8 pg, and 1.6 pg) were amplified by primers P1 and P2 under identical PCR conditions. (F) Chromatin isolated from TNF-treated or control cells fixed with formaldehyde was subjected to NcoI digestion as indicated. Following deproteination, the DNA was purified and analyzed by PCR using the indicated primers. Because the ligation step of the 3C assay was not performed, P1/P2, P1/P6, and P2/P5 PCR products can only be observed when NcoI was omitted from the reaction, indicating that the chromatin is accessible to restriction digestion even in the absence of TNF.

    Journal: Molecular and Cellular Biology

    Article Title: Mechanism of Action of a Distal NF-?B-Dependent Enhancer

    doi: 10.1128/MCB.00271-06

    Figure Lengend Snippet: An interaction between the distal and proximal regulatory regions of the MCP - 1 gene forms following TNF stimulation. (A) A schematic representation of the 3C assay and the architecture of the MCP - 1 promoter with cis regulatory sites with selected restriction sites and primer positions (P1 through P4) is shown. In the 3C assay, cells were fixed with formaldehyde and chromatin was isolated and digested with NcoI. Following inactivation of the restriction enzyme, the sample was diluted and subjected to DNA ligation such that only intramolecular ligations would be preferred. The cross-links were then reversed, and the DNA was purified. PCR primers P1 and P2 were used to detect the formation of the novel ligated 3C product. Primers P3 and P4 were used to verify that similar levels of MCP - 1 DNA were present in the assay and that the DNA was amplifiable. Products from the 3C assay were examined on agarose gels stained with ethidium bromide. (B) NIH 3T3 cells treated in the absence (−) (lanes 1 to 4) or presence (+) (lanes 5 to 8) of TNF for 2 h were processed in the above 3C assay with and without formaldehyde (CH 2 O) or DNA ligase, as indicated, to control for specificity of the protocol. M, DNA marker. (C) Purified mouse genomic DNA was processed in the 3C assay and demonstrates that the unique PCR product cannot form with naked DNA templates. (D) KpnI cleaves the PCR product generated by the 3C assay, producing the anticipated DNA fragments as shown on the agarose gel. (E) PCR amplification of the 3C product and the non-3C product display similar relative efficiencies. PCR products generated from undigested genomic DNA (1,207 bp) and DNA from a 3C assay (713 bp) were quantitated by fluorometry and used as templates in PCRs to determine if there were major differences in their PCR efficiency. DNA templates (40 pg, 8 pg, and 1.6 pg) were amplified by primers P1 and P2 under identical PCR conditions. (F) Chromatin isolated from TNF-treated or control cells fixed with formaldehyde was subjected to NcoI digestion as indicated. Following deproteination, the DNA was purified and analyzed by PCR using the indicated primers. Because the ligation step of the 3C assay was not performed, P1/P2, P1/P6, and P2/P5 PCR products can only be observed when NcoI was omitted from the reaction, indicating that the chromatin is accessible to restriction digestion even in the absence of TNF.

    Article Snippet: The cross-linked DNA was digested overnight with 500 to 800 units NcoI restriction enzyme (New England Biolabs, Inc.).

    Techniques: Isolation, DNA Ligation, Purification, Polymerase Chain Reaction, Staining, Marker, Generated, Agarose Gel Electrophoresis, Amplification, Ligation