restriction endonucleases bcci  (New England Biolabs)


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    Structured Review

    New England Biolabs restriction endonucleases bcci
    <t>pET-BccI</t> untreated and digested. 1 : <t>DNA</t> ladder. 2 : pET-BccI untreated. 3 : pET-BccI digested with BccI. 4 , 5 , 6 : pET-BccI digested with EcoRI, BamHI and HindIII, respectively.
    Restriction Endonucleases Bcci, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction endonucleases bcci/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    restriction endonucleases bcci - by Bioz Stars, 2021-04
    86/100 stars

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    1) Product Images from "TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression"

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0186568

    pET-BccI untreated and digested. 1 : DNA ladder. 2 : pET-BccI untreated. 3 : pET-BccI digested with BccI. 4 , 5 , 6 : pET-BccI digested with EcoRI, BamHI and HindIII, respectively.
    Figure Legend Snippet: pET-BccI untreated and digested. 1 : DNA ladder. 2 : pET-BccI untreated. 3 : pET-BccI digested with BccI. 4 , 5 , 6 : pET-BccI digested with EcoRI, BamHI and HindIII, respectively.

    Techniques Used: Positron Emission Tomography

    The procedure of TA-GC cloning. The target protein-coding gene is initially amplified using PCR and the PCR product is treated with T4 DNA polymerase. In parallel, the vector is digested with BccI. Afterwards, a ligation reaction between the purified linearized vector and the protein-coding gene is set up followed by transformation in high efficiency chemocompetent E . coli cells.
    Figure Legend Snippet: The procedure of TA-GC cloning. The target protein-coding gene is initially amplified using PCR and the PCR product is treated with T4 DNA polymerase. In parallel, the vector is digested with BccI. Afterwards, a ligation reaction between the purified linearized vector and the protein-coding gene is set up followed by transformation in high efficiency chemocompetent E . coli cells.

    Techniques Used: Clone Assay, Amplification, Polymerase Chain Reaction, Plasmid Preparation, Ligation, Purification, Transformation Assay

    Screening for E . coli colonies transformed with recombinant pET-BccI. Plasmid preparations from cultures inoculated with transformed colonies were digested with BamHI and subjected to electrophoresis. A ) For BRP recombinants pET-BccI screening, the presence of a 283 bp DNA band indicated there was no recombination (A3, A7, A9), whereas the presence of a 636 bp insert DNA band showed a successful recombination (A2, A4, A5, A6, A8). B ) For CAT recombinants pET-BccI screening, again the presence of a 283 bp DNA band indicated there was no recombination (B2, B5, B6, B8), whereas the presence of a 924 bp insert DNA band showed a successful recombination (B3, B4, B7, B9).
    Figure Legend Snippet: Screening for E . coli colonies transformed with recombinant pET-BccI. Plasmid preparations from cultures inoculated with transformed colonies were digested with BamHI and subjected to electrophoresis. A ) For BRP recombinants pET-BccI screening, the presence of a 283 bp DNA band indicated there was no recombination (A3, A7, A9), whereas the presence of a 636 bp insert DNA band showed a successful recombination (A2, A4, A5, A6, A8). B ) For CAT recombinants pET-BccI screening, again the presence of a 283 bp DNA band indicated there was no recombination (B2, B5, B6, B8), whereas the presence of a 924 bp insert DNA band showed a successful recombination (B3, B4, B7, B9).

    Techniques Used: Transformation Assay, Recombinant, Positron Emission Tomography, Plasmid Preparation, Electrophoresis

    TA-GC cloning. A : BccI recognizes the sequence CCATC at the cloning site of pET-BccI and cuts out at the recognition site as indicated. B : after digestion with BccI. C : the protein-coding gene (starting with an ATG codon and always having a 3΄ glycine-coding GGC codon) after PCR amplification, and D : after treatment with T4 DNA polymerase in the presence of dATP and dGTP. The 5΄-Α and G overhangs of the protein-coding gene created after incubation with the T4 DNA polymerase are complementary to the 5΄-T and C overhangs of the pET-BccI vector after digestion with BccI.
    Figure Legend Snippet: TA-GC cloning. A : BccI recognizes the sequence CCATC at the cloning site of pET-BccI and cuts out at the recognition site as indicated. B : after digestion with BccI. C : the protein-coding gene (starting with an ATG codon and always having a 3΄ glycine-coding GGC codon) after PCR amplification, and D : after treatment with T4 DNA polymerase in the presence of dATP and dGTP. The 5΄-Α and G overhangs of the protein-coding gene created after incubation with the T4 DNA polymerase are complementary to the 5΄-T and C overhangs of the pET-BccI vector after digestion with BccI.

    Techniques Used: Clone Assay, Sequencing, Positron Emission Tomography, Polymerase Chain Reaction, Amplification, Incubation, Plasmid Preparation

    The novel protein-expression vector pET-BccI. The pET-26b (+) derived plasmid has a pBR322 origin of replication, which together with the ROP protein regulates the plasmid copy number per bacterial cell. The kanamycin resistance gene enables positive selection of the transformed E . coli cells in the presence of kanamycin. BamHI, EcoRI and HindIII recognition sites, flanking both sites of the T7 promoter, cloning site and T7 terminator cassette, facilitate the screening of the transformed colonies for the recombinant transformants. The cloning site of pET-BccI, composed of two adjacent reverse BccI recognition sites, provides single 5΄-T and C overhangs after digestion with BccI, which are suitable for the ligation of DNA molecules with complementary edges.
    Figure Legend Snippet: The novel protein-expression vector pET-BccI. The pET-26b (+) derived plasmid has a pBR322 origin of replication, which together with the ROP protein regulates the plasmid copy number per bacterial cell. The kanamycin resistance gene enables positive selection of the transformed E . coli cells in the presence of kanamycin. BamHI, EcoRI and HindIII recognition sites, flanking both sites of the T7 promoter, cloning site and T7 terminator cassette, facilitate the screening of the transformed colonies for the recombinant transformants. The cloning site of pET-BccI, composed of two adjacent reverse BccI recognition sites, provides single 5΄-T and C overhangs after digestion with BccI, which are suitable for the ligation of DNA molecules with complementary edges.

    Techniques Used: Expressing, Plasmid Preparation, Positron Emission Tomography, Derivative Assay, Selection, Transformation Assay, Clone Assay, Recombinant, Ligation

    Related Articles

    Purification:

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression
    Article Snippet: Plasmid DNA was prepared using the NucleoSpin Plasmid purification kit (Macherey-Nagel), according to the manufacturer’s protocol. .. The purified pET-BccI batches were analyzed through the digestion of 1 μg plasmid DNA with 1 unit of the restriction endonucleases BccI, EcoRI, BamHI and HindIII (New England BioLabs), for 1 hour, at 37°C in the NEB buffers: CutSmart, EcoRI, 3.1 and 2.1, respectively. .. Cloning using TA-GC method Initially the protein coding gene is amplified using primers that yield PCR product flanked by an ATG methionine codon and a GGC glycine codon , or as discussed in the aim of the study.

    Positron Emission Tomography:

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression
    Article Snippet: Plasmid DNA was prepared using the NucleoSpin Plasmid purification kit (Macherey-Nagel), according to the manufacturer’s protocol. .. The purified pET-BccI batches were analyzed through the digestion of 1 μg plasmid DNA with 1 unit of the restriction endonucleases BccI, EcoRI, BamHI and HindIII (New England BioLabs), for 1 hour, at 37°C in the NEB buffers: CutSmart, EcoRI, 3.1 and 2.1, respectively. .. Cloning using TA-GC method Initially the protein coding gene is amplified using primers that yield PCR product flanked by an ATG methionine codon and a GGC glycine codon , or as discussed in the aim of the study.

    Plasmid Preparation:

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression
    Article Snippet: Plasmid DNA was prepared using the NucleoSpin Plasmid purification kit (Macherey-Nagel), according to the manufacturer’s protocol. .. The purified pET-BccI batches were analyzed through the digestion of 1 μg plasmid DNA with 1 unit of the restriction endonucleases BccI, EcoRI, BamHI and HindIII (New England BioLabs), for 1 hour, at 37°C in the NEB buffers: CutSmart, EcoRI, 3.1 and 2.1, respectively. .. Cloning using TA-GC method Initially the protein coding gene is amplified using primers that yield PCR product flanked by an ATG methionine codon and a GGC glycine codon , or as discussed in the aim of the study.

    Labeling:

    Article Title: Mutation in the RmβAOR gene is associated with amitraz resistance in the cattle tick Rhipicephalus microplus
    Article Snippet: .. Genotyping was carried out by digesting the fluorescently labeled PCR product with the restriction endonuclease BCCI (New England Biolabs) using 2.5 U of enzyme, 1× NE Buffer 1 supplemented with 100 μg/mL BSA at 37 °C, overnight. ..

    Polymerase Chain Reaction:

    Article Title: Mutation in the RmβAOR gene is associated with amitraz resistance in the cattle tick Rhipicephalus microplus
    Article Snippet: .. Genotyping was carried out by digesting the fluorescently labeled PCR product with the restriction endonuclease BCCI (New England Biolabs) using 2.5 U of enzyme, 1× NE Buffer 1 supplemented with 100 μg/mL BSA at 37 °C, overnight. ..

    Sequencing:

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression
    Article Snippet: After PCR amplification and a short T4 DNA polymerase treatment, protein-coding genes obtain single 5- A and G overhangs, complementary to the 5΄- T and C overhangs of a linearized vector, like pET-BccI, after digestion with the restriction endonuclease BccI ( ). .. BccI is a category B restriction endonuclease which recognizes the sequence CCATC and cuts out of the recognition site, 4 bases towards the 3’ direction at the main DNA strand and 5 bases towards the 5΄ direction at the complementary strand, thus creating single 5΄ nucleotide overhangs (New England BioLabs). .. Vectors like pET-BccI can be created from existing plasmids by replacing the original cloning sites with the cloning site of pET-BccI, while omitting the BccI recognition sites apart from those of the cloning site.

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    New England Biolabs restriction endonuclease bcci
    Restriction Endonuclease Bcci, 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/restriction endonuclease bcci/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    restriction endonuclease bcci - by Bioz Stars, 2021-04
    99/100 stars
      Buy from Supplier

    86
    New England Biolabs restriction endonucleases bcci
    <t>pET-BccI</t> untreated and digested. 1 : <t>DNA</t> ladder. 2 : pET-BccI untreated. 3 : pET-BccI digested with BccI. 4 , 5 , 6 : pET-BccI digested with EcoRI, BamHI and HindIII, respectively.
    Restriction Endonucleases Bcci, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction endonucleases bcci/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    restriction endonucleases bcci - by Bioz Stars, 2021-04
    86/100 stars
      Buy from Supplier

    Image Search Results


    pET-BccI untreated and digested. 1 : DNA ladder. 2 : pET-BccI untreated. 3 : pET-BccI digested with BccI. 4 , 5 , 6 : pET-BccI digested with EcoRI, BamHI and HindIII, respectively.

    Journal: PLoS ONE

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression

    doi: 10.1371/journal.pone.0186568

    Figure Lengend Snippet: pET-BccI untreated and digested. 1 : DNA ladder. 2 : pET-BccI untreated. 3 : pET-BccI digested with BccI. 4 , 5 , 6 : pET-BccI digested with EcoRI, BamHI and HindIII, respectively.

    Article Snippet: The purified pET-BccI batches were analyzed through the digestion of 1 μg plasmid DNA with 1 unit of the restriction endonucleases BccI, EcoRI, BamHI and HindIII (New England BioLabs), for 1 hour, at 37°C in the NEB buffers: CutSmart, EcoRI, 3.1 and 2.1, respectively.

    Techniques: Positron Emission Tomography

    The procedure of TA-GC cloning. The target protein-coding gene is initially amplified using PCR and the PCR product is treated with T4 DNA polymerase. In parallel, the vector is digested with BccI. Afterwards, a ligation reaction between the purified linearized vector and the protein-coding gene is set up followed by transformation in high efficiency chemocompetent E . coli cells.

    Journal: PLoS ONE

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression

    doi: 10.1371/journal.pone.0186568

    Figure Lengend Snippet: The procedure of TA-GC cloning. The target protein-coding gene is initially amplified using PCR and the PCR product is treated with T4 DNA polymerase. In parallel, the vector is digested with BccI. Afterwards, a ligation reaction between the purified linearized vector and the protein-coding gene is set up followed by transformation in high efficiency chemocompetent E . coli cells.

    Article Snippet: The purified pET-BccI batches were analyzed through the digestion of 1 μg plasmid DNA with 1 unit of the restriction endonucleases BccI, EcoRI, BamHI and HindIII (New England BioLabs), for 1 hour, at 37°C in the NEB buffers: CutSmart, EcoRI, 3.1 and 2.1, respectively.

    Techniques: Clone Assay, Amplification, Polymerase Chain Reaction, Plasmid Preparation, Ligation, Purification, Transformation Assay

    Screening for E . coli colonies transformed with recombinant pET-BccI. Plasmid preparations from cultures inoculated with transformed colonies were digested with BamHI and subjected to electrophoresis. A ) For BRP recombinants pET-BccI screening, the presence of a 283 bp DNA band indicated there was no recombination (A3, A7, A9), whereas the presence of a 636 bp insert DNA band showed a successful recombination (A2, A4, A5, A6, A8). B ) For CAT recombinants pET-BccI screening, again the presence of a 283 bp DNA band indicated there was no recombination (B2, B5, B6, B8), whereas the presence of a 924 bp insert DNA band showed a successful recombination (B3, B4, B7, B9).

    Journal: PLoS ONE

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression

    doi: 10.1371/journal.pone.0186568

    Figure Lengend Snippet: Screening for E . coli colonies transformed with recombinant pET-BccI. Plasmid preparations from cultures inoculated with transformed colonies were digested with BamHI and subjected to electrophoresis. A ) For BRP recombinants pET-BccI screening, the presence of a 283 bp DNA band indicated there was no recombination (A3, A7, A9), whereas the presence of a 636 bp insert DNA band showed a successful recombination (A2, A4, A5, A6, A8). B ) For CAT recombinants pET-BccI screening, again the presence of a 283 bp DNA band indicated there was no recombination (B2, B5, B6, B8), whereas the presence of a 924 bp insert DNA band showed a successful recombination (B3, B4, B7, B9).

    Article Snippet: The purified pET-BccI batches were analyzed through the digestion of 1 μg plasmid DNA with 1 unit of the restriction endonucleases BccI, EcoRI, BamHI and HindIII (New England BioLabs), for 1 hour, at 37°C in the NEB buffers: CutSmart, EcoRI, 3.1 and 2.1, respectively.

    Techniques: Transformation Assay, Recombinant, Positron Emission Tomography, Plasmid Preparation, Electrophoresis

    TA-GC cloning. A : BccI recognizes the sequence CCATC at the cloning site of pET-BccI and cuts out at the recognition site as indicated. B : after digestion with BccI. C : the protein-coding gene (starting with an ATG codon and always having a 3΄ glycine-coding GGC codon) after PCR amplification, and D : after treatment with T4 DNA polymerase in the presence of dATP and dGTP. The 5΄-Α and G overhangs of the protein-coding gene created after incubation with the T4 DNA polymerase are complementary to the 5΄-T and C overhangs of the pET-BccI vector after digestion with BccI.

    Journal: PLoS ONE

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression

    doi: 10.1371/journal.pone.0186568

    Figure Lengend Snippet: TA-GC cloning. A : BccI recognizes the sequence CCATC at the cloning site of pET-BccI and cuts out at the recognition site as indicated. B : after digestion with BccI. C : the protein-coding gene (starting with an ATG codon and always having a 3΄ glycine-coding GGC codon) after PCR amplification, and D : after treatment with T4 DNA polymerase in the presence of dATP and dGTP. The 5΄-Α and G overhangs of the protein-coding gene created after incubation with the T4 DNA polymerase are complementary to the 5΄-T and C overhangs of the pET-BccI vector after digestion with BccI.

    Article Snippet: The purified pET-BccI batches were analyzed through the digestion of 1 μg plasmid DNA with 1 unit of the restriction endonucleases BccI, EcoRI, BamHI and HindIII (New England BioLabs), for 1 hour, at 37°C in the NEB buffers: CutSmart, EcoRI, 3.1 and 2.1, respectively.

    Techniques: Clone Assay, Sequencing, Positron Emission Tomography, Polymerase Chain Reaction, Amplification, Incubation, Plasmid Preparation

    The novel protein-expression vector pET-BccI. The pET-26b (+) derived plasmid has a pBR322 origin of replication, which together with the ROP protein regulates the plasmid copy number per bacterial cell. The kanamycin resistance gene enables positive selection of the transformed E . coli cells in the presence of kanamycin. BamHI, EcoRI and HindIII recognition sites, flanking both sites of the T7 promoter, cloning site and T7 terminator cassette, facilitate the screening of the transformed colonies for the recombinant transformants. The cloning site of pET-BccI, composed of two adjacent reverse BccI recognition sites, provides single 5΄-T and C overhangs after digestion with BccI, which are suitable for the ligation of DNA molecules with complementary edges.

    Journal: PLoS ONE

    Article Title: TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression

    doi: 10.1371/journal.pone.0186568

    Figure Lengend Snippet: The novel protein-expression vector pET-BccI. The pET-26b (+) derived plasmid has a pBR322 origin of replication, which together with the ROP protein regulates the plasmid copy number per bacterial cell. The kanamycin resistance gene enables positive selection of the transformed E . coli cells in the presence of kanamycin. BamHI, EcoRI and HindIII recognition sites, flanking both sites of the T7 promoter, cloning site and T7 terminator cassette, facilitate the screening of the transformed colonies for the recombinant transformants. The cloning site of pET-BccI, composed of two adjacent reverse BccI recognition sites, provides single 5΄-T and C overhangs after digestion with BccI, which are suitable for the ligation of DNA molecules with complementary edges.

    Article Snippet: The purified pET-BccI batches were analyzed through the digestion of 1 μg plasmid DNA with 1 unit of the restriction endonucleases BccI, EcoRI, BamHI and HindIII (New England BioLabs), for 1 hour, at 37°C in the NEB buffers: CutSmart, EcoRI, 3.1 and 2.1, respectively.

    Techniques: Expressing, Plasmid Preparation, Positron Emission Tomography, Derivative Assay, Selection, Transformation Assay, Clone Assay, Recombinant, Ligation