flanking enzymes ncoi  (New England Biolabs)


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  • 99
    Name:
    NcoI
    Description:
    NcoI 5 000 units
    Catalog Number:
    r0193l
    Price:
    261
    Size:
    5 000 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs flanking enzymes ncoi
    NcoI
    NcoI 5 000 units
    https://www.bioz.com/result/flanking enzymes ncoi/product/New England Biolabs
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    flanking enzymes ncoi - by Bioz Stars, 2020-07
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Fluorescence-based methods for measuring target interference by CRISPR-Cas systems
    Article Snippet: .. T4 polynucleotide kinase (PNK), NotI, NcoI, T4 DNA ligase and accompanying buffers purchased from New England Biolabs 100 mM ATP 100 µM CRISPR target oligonucleotides (designed as in ) and pACYC-GFP Gel purification kit (e.g. Qiagen QIAquick Gel Extraction kit or Promega Wizard SV Gel and PCR Clean-Up System) One Shot TOP10 Competent Cells (Thermo-Fisher) or similar cloning E. coli strain Miniprep kit (Qiagen or Promega) ..

    Agarose Gel Electrophoresis:

    Article Title: β-nicotinamide mononucleotide (NMN) production in Escherichia coli
    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. ). .. Resulted bicistronic vector pET28a-baPrs-hdNadV was deposited at Addgene under the ID #91950.

    Polyacrylamide Gel Electrophoresis:

    Article Title: A Quantitative Assay for Assessing the Effects of DNA Lesions on Transcription
    Article Snippet: .. PAGE analysis In order to analyze the transcription products of S -cdAand S -cdG using PAGE, a portion of the above RT-PCR fragments was treated with 10 U NcoI and 1 U shrimp alkaline phosphatase in 10 μL NEB buffer 3 at 37°C for 30 min, followed by heating at 80°C for 20 min to deactivate the phosphatase. ..

    Ligation:

    Article Title: Chloride-Inducible Expression Vector for Delivery of Antimicrobial Peptides Targeting Antibiotic-Resistant Enterococcus faecium
    Article Snippet: .. The PCR fragment and plasmid pNZ8048 were both digested with restriction enzymes BglII and NcoI (New England BioLabs) and ligated at 16°C for 16 h. The resulting ligation was then transformed into electrocompetent E. coli MC1061 F′ (Lucigen). .. Successful transformants were identified by colony PCR using the primers pNZ8048-F (5′-GCCCCGTTAGTTGAAGAAGG-3′) and pNZ8048-R (5′-CAATTGAACGTTTCAAGCCTTGG-3′) and further verified by sequencing analysis.

    Article Title: β-nicotinamide mononucleotide (NMN) production in Escherichia coli
    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. ). .. Resulted bicistronic vector pET28a-baPrs-hdNadV was deposited at Addgene under the ID #91950.

    Construct:

    Article Title: β-nicotinamide mononucleotide (NMN) production in Escherichia coli
    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. ). .. Resulted bicistronic vector pET28a-baPrs-hdNadV was deposited at Addgene under the ID #91950.

    Purification:

    Article Title: β-nicotinamide mononucleotide (NMN) production in Escherichia coli
    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. ). .. Resulted bicistronic vector pET28a-baPrs-hdNadV was deposited at Addgene under the ID #91950.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: A Quantitative Assay for Assessing the Effects of DNA Lesions on Transcription
    Article Snippet: .. PAGE analysis In order to analyze the transcription products of S -cdAand S -cdG using PAGE, a portion of the above RT-PCR fragments was treated with 10 U NcoI and 1 U shrimp alkaline phosphatase in 10 μL NEB buffer 3 at 37°C for 30 min, followed by heating at 80°C for 20 min to deactivate the phosphatase. ..

    Article Title: A Quantitative Assay for Assessing the Effects of DNA Lesions on Transcription
    Article Snippet: .. LC-MS/MS analysis In order to identify the transcription products of S -cdA and S -cdG using LC-MS/MS, RT-PCR products were treated with 50 U NcoI and 20 U shrimp alkaline phosphatase in 250 μL NEB buffer 3 at 37°C for 2 h, followed by heating at 80°C for 20 min. To the resulting solution was added 50 U of AseI, and the reaction mixture was incubated at 37°C for 1 h, followed by extraction once with phenol/chloroform/isoamyl alcohol (25:24:1, v/v). .. The aqueous portion was dried with Speed-vac, desalted with HPLC and dissolved in 12 μL water.

    Incubation:

    Article Title: A Quantitative Assay for Assessing the Effects of DNA Lesions on Transcription
    Article Snippet: .. LC-MS/MS analysis In order to identify the transcription products of S -cdA and S -cdG using LC-MS/MS, RT-PCR products were treated with 50 U NcoI and 20 U shrimp alkaline phosphatase in 250 μL NEB buffer 3 at 37°C for 2 h, followed by heating at 80°C for 20 min. To the resulting solution was added 50 U of AseI, and the reaction mixture was incubated at 37°C for 1 h, followed by extraction once with phenol/chloroform/isoamyl alcohol (25:24:1, v/v). .. The aqueous portion was dried with Speed-vac, desalted with HPLC and dissolved in 12 μL water.

    Polymerase Chain Reaction:

    Article Title: Fluorescence-based methods for measuring target interference by CRISPR-Cas systems
    Article Snippet: .. T4 polynucleotide kinase (PNK), NotI, NcoI, T4 DNA ligase and accompanying buffers purchased from New England Biolabs 100 mM ATP 100 µM CRISPR target oligonucleotides (designed as in ) and pACYC-GFP Gel purification kit (e.g. Qiagen QIAquick Gel Extraction kit or Promega Wizard SV Gel and PCR Clean-Up System) One Shot TOP10 Competent Cells (Thermo-Fisher) or similar cloning E. coli strain Miniprep kit (Qiagen or Promega) ..

    Article Title: Chloride-Inducible Expression Vector for Delivery of Antimicrobial Peptides Targeting Antibiotic-Resistant Enterococcus faecium
    Article Snippet: .. The PCR fragment and plasmid pNZ8048 were both digested with restriction enzymes BglII and NcoI (New England BioLabs) and ligated at 16°C for 16 h. The resulting ligation was then transformed into electrocompetent E. coli MC1061 F′ (Lucigen). .. Successful transformants were identified by colony PCR using the primers pNZ8048-F (5′-GCCCCGTTAGTTGAAGAAGG-3′) and pNZ8048-R (5′-CAATTGAACGTTTCAAGCCTTGG-3′) and further verified by sequencing analysis.

    Article Title: β-nicotinamide mononucleotide (NMN) production in Escherichia coli
    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. ). .. Resulted bicistronic vector pET28a-baPrs-hdNadV was deposited at Addgene under the ID #91950.

    Expressing:

    Article Title: β-nicotinamide mononucleotide (NMN) production in Escherichia coli
    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. ). .. Resulted bicistronic vector pET28a-baPrs-hdNadV was deposited at Addgene under the ID #91950.

    CRISPR:

    Article Title: Fluorescence-based methods for measuring target interference by CRISPR-Cas systems
    Article Snippet: .. T4 polynucleotide kinase (PNK), NotI, NcoI, T4 DNA ligase and accompanying buffers purchased from New England Biolabs 100 mM ATP 100 µM CRISPR target oligonucleotides (designed as in ) and pACYC-GFP Gel purification kit (e.g. Qiagen QIAquick Gel Extraction kit or Promega Wizard SV Gel and PCR Clean-Up System) One Shot TOP10 Competent Cells (Thermo-Fisher) or similar cloning E. coli strain Miniprep kit (Qiagen or Promega) ..

    Sequencing:

    Article Title: β-nicotinamide mononucleotide (NMN) production in Escherichia coli
    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. ). .. Resulted bicistronic vector pET28a-baPrs-hdNadV was deposited at Addgene under the ID #91950.

    Gel Extraction:

    Article Title: Fluorescence-based methods for measuring target interference by CRISPR-Cas systems
    Article Snippet: .. T4 polynucleotide kinase (PNK), NotI, NcoI, T4 DNA ligase and accompanying buffers purchased from New England Biolabs 100 mM ATP 100 µM CRISPR target oligonucleotides (designed as in ) and pACYC-GFP Gel purification kit (e.g. Qiagen QIAquick Gel Extraction kit or Promega Wizard SV Gel and PCR Clean-Up System) One Shot TOP10 Competent Cells (Thermo-Fisher) or similar cloning E. coli strain Miniprep kit (Qiagen or Promega) ..

    Transformation Assay:

    Article Title: Chloride-Inducible Expression Vector for Delivery of Antimicrobial Peptides Targeting Antibiotic-Resistant Enterococcus faecium
    Article Snippet: .. The PCR fragment and plasmid pNZ8048 were both digested with restriction enzymes BglII and NcoI (New England BioLabs) and ligated at 16°C for 16 h. The resulting ligation was then transformed into electrocompetent E. coli MC1061 F′ (Lucigen). .. Successful transformants were identified by colony PCR using the primers pNZ8048-F (5′-GCCCCGTTAGTTGAAGAAGG-3′) and pNZ8048-R (5′-CAATTGAACGTTTCAAGCCTTGG-3′) and further verified by sequencing analysis.

    Plasmid Preparation:

    Article Title: Codon Usage Optimization and Construction of Plasmid Encoding Iranian Human Papillomavirus Type 16 E7 Oncogene for Lactococcus Lactis Subsp. Cremoris MG1363
    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). .. The resultant DNA fragment was ligated into the NcoI/SacI site of pNZ8148 shuttle vector (MoBiTec, Germany).

    Article Title: Chloride-Inducible Expression Vector for Delivery of Antimicrobial Peptides Targeting Antibiotic-Resistant Enterococcus faecium
    Article Snippet: .. The PCR fragment and plasmid pNZ8048 were both digested with restriction enzymes BglII and NcoI (New England BioLabs) and ligated at 16°C for 16 h. The resulting ligation was then transformed into electrocompetent E. coli MC1061 F′ (Lucigen). .. Successful transformants were identified by colony PCR using the primers pNZ8048-F (5′-GCCCCGTTAGTTGAAGAAGG-3′) and pNZ8048-R (5′-CAATTGAACGTTTCAAGCCTTGG-3′) and further verified by sequencing analysis.

    Article Title: β-nicotinamide mononucleotide (NMN) production in Escherichia coli
    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. ). .. Resulted bicistronic vector pET28a-baPrs-hdNadV was deposited at Addgene under the ID #91950.

    Gel Purification:

    Article Title: Fluorescence-based methods for measuring target interference by CRISPR-Cas systems
    Article Snippet: .. T4 polynucleotide kinase (PNK), NotI, NcoI, T4 DNA ligase and accompanying buffers purchased from New England Biolabs 100 mM ATP 100 µM CRISPR target oligonucleotides (designed as in ) and pACYC-GFP Gel purification kit (e.g. Qiagen QIAquick Gel Extraction kit or Promega Wizard SV Gel and PCR Clean-Up System) One Shot TOP10 Competent Cells (Thermo-Fisher) or similar cloning E. coli strain Miniprep kit (Qiagen or Promega) ..

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  • 99
    New England Biolabs flanking enzymes ncoi
    Flanking Enzymes Ncoi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flanking enzymes ncoi/product/New England Biolabs
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    flanking enzymes ncoi - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

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