restriction endonucleases bcci  (New England Biolabs)


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    Name:
    BccI
    Description:
    BccI 5 000 units
    Catalog Number:
    r0704l
    Price:
    282
    Size:
    5 000 units
    Category:
    Restriction Enzymes
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    New England Biolabs restriction endonucleases bcci
    BccI
    BccI 5 000 units
    https://www.bioz.com/result/restriction endonucleases bcci/product/New England Biolabs
    Average 94 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    restriction endonucleases bcci - by Bioz Stars, 2020-09
    94/100 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

    2) Product Images from "Kinesin Family Member 6 (kif6) Is Necessary for Spine Development in Zebrafish"

    Article Title: Kinesin Family Member 6 (kif6) Is Necessary for Spine Development in Zebrafish

    Journal: Developmental dynamics : an official publication of the American Association of Anatomists

    doi: 10.1002/dvdy.24208

    Design and efficiency of kif6 TALENs. A: Genomic sequence showing left and right TALENs that were designed in the second exon of kif6 near the gw326 mutant allele. The TALEN pair flank a BccI restriction site. B: BccI restriction digest of PCR amplified
    Figure Legend Snippet: Design and efficiency of kif6 TALENs. A: Genomic sequence showing left and right TALENs that were designed in the second exon of kif6 near the gw326 mutant allele. The TALEN pair flank a BccI restriction site. B: BccI restriction digest of PCR amplified

    Techniques Used: TALENs, Sequencing, Mutagenesis, Polymerase Chain Reaction, Amplification

    3) Product Images from "Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform"

    Article Title: Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform

    Journal: Biosensors & bioelectronics

    doi: 10.1016/j.bios.2019.111678

    Sequence alignment, designed primers and analytical specificity for the detection of P. falciparum . (A) Illustration of the gene kelch 13 highlighting the location of the LAMP-PfK13 amplicon. (B) Alignment of all human-infective Plasmodium species with P. falciparum as reference sequence. Mismatches in the alignment are displayed as AGTC whereas matched nucleotides are represented as dots. The LAMP-PfK13 primers are illustrated on top of the alignment. (C) . (D) Results showing specific amplification of P. falciparum . The restriction enzyme used for this experiment is BccI (# R0704S, New England BioLabs). The cutting point is illustrated with an arrow and the binding region is orange shadowed in (B). M denotes 100 bp DNA ladder (#10488058, Invitrogen). Acronyms are described in (B). (E) ] used as control for the detection of all Plasmodium species. TTP values are displayed in minutes and labelled.
    Figure Legend Snippet: Sequence alignment, designed primers and analytical specificity for the detection of P. falciparum . (A) Illustration of the gene kelch 13 highlighting the location of the LAMP-PfK13 amplicon. (B) Alignment of all human-infective Plasmodium species with P. falciparum as reference sequence. Mismatches in the alignment are displayed as AGTC whereas matched nucleotides are represented as dots. The LAMP-PfK13 primers are illustrated on top of the alignment. (C) . (D) Results showing specific amplification of P. falciparum . The restriction enzyme used for this experiment is BccI (# R0704S, New England BioLabs). The cutting point is illustrated with an arrow and the binding region is orange shadowed in (B). M denotes 100 bp DNA ladder (#10488058, Invitrogen). Acronyms are described in (B). (E) ] used as control for the detection of all Plasmodium species. TTP values are displayed in minutes and labelled.

    Techniques Used: Sequencing, Amplification, Binding Assay

    4) Product Images from "Genetic Polymorphism and Expression of CXCR4 in Breast Cancer"

    Article Title: Genetic Polymorphism and Expression of CXCR4 in Breast Cancer

    Journal: Analytical cellular pathology (Amsterdam)

    doi: 10.1155/2015/289510

    CXCR4 rs2228014 (C/T) genetic polymorphism. (a) Electrophoretic profile of rs2228014 (C/T). BccI restriction enzyme was used for 4 h at 37°C. Polyacrylamide gel 10% stained with silver nitrate. Lane 1, Ladder DNA fragment marker of 100 bp; Lane 2, PCR product of 236 pb; Lane 3, wild-type homozygous genotype of 133 pb and 103 pb (CC); Lane 4, heterozygous genotype of 236 pb, 133 pb, and 103 pb (CT); Lane 5, blank reaction or negative control (reaction without DNA). (b) Genotype distribution for CXCR4 rs2228014 in breast cancer patients.
    Figure Legend Snippet: CXCR4 rs2228014 (C/T) genetic polymorphism. (a) Electrophoretic profile of rs2228014 (C/T). BccI restriction enzyme was used for 4 h at 37°C. Polyacrylamide gel 10% stained with silver nitrate. Lane 1, Ladder DNA fragment marker of 100 bp; Lane 2, PCR product of 236 pb; Lane 3, wild-type homozygous genotype of 133 pb and 103 pb (CC); Lane 4, heterozygous genotype of 236 pb, 133 pb, and 103 pb (CT); Lane 5, blank reaction or negative control (reaction without DNA). (b) Genotype distribution for CXCR4 rs2228014 in breast cancer patients.

    Techniques Used: Staining, Marker, Polymerase Chain Reaction, Negative Control

    5) Product Images from "Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform"

    Article Title: Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform

    Journal: bioRxiv

    doi: 10.1101/638221

    Sequence alignment, primer design and analytical specificity for the detection of P. falciparum . (A) Illustration of the gene kelch 13 showing in black the region selected for primer design. (B) Alignment of all human-infective Plasmodium species with P. falciparum as reference sequence. Mismatches in the alignment are displayed as AGTC whereas matched nucleotides are represented as dots. Primers are illustrated on top of the alignment. (C) Primer location in LAMP amplicon (5’ to 3’ direction). Sequences of the primers can be found in table S1. (D) Results showing specific amplification of P. falciparum DNA samples from clinical isolates as well as gel electrophoresis of the amplified products to confirm specificity. Digested amplification products are shown in fig. S2. The restriction enzyme used for this experiment is BccI (# R0704S, New England BioLabs). The cutting point is illustrated with an arrow and the binding region is orange shadowed in (B). M denotes 100 bp DNA ladder (#10488058, Invitrogen). Samples are described in (B). (E) Pan-primer set [ 21 ] used as reference for the detection of all Plasmodium species. TTP values are displayed in minutes and labelled.
    Figure Legend Snippet: Sequence alignment, primer design and analytical specificity for the detection of P. falciparum . (A) Illustration of the gene kelch 13 showing in black the region selected for primer design. (B) Alignment of all human-infective Plasmodium species with P. falciparum as reference sequence. Mismatches in the alignment are displayed as AGTC whereas matched nucleotides are represented as dots. Primers are illustrated on top of the alignment. (C) Primer location in LAMP amplicon (5’ to 3’ direction). Sequences of the primers can be found in table S1. (D) Results showing specific amplification of P. falciparum DNA samples from clinical isolates as well as gel electrophoresis of the amplified products to confirm specificity. Digested amplification products are shown in fig. S2. The restriction enzyme used for this experiment is BccI (# R0704S, New England BioLabs). The cutting point is illustrated with an arrow and the binding region is orange shadowed in (B). M denotes 100 bp DNA ladder (#10488058, Invitrogen). Samples are described in (B). (E) Pan-primer set [ 21 ] used as reference for the detection of all Plasmodium species. TTP values are displayed in minutes and labelled.

    Techniques Used: Sequencing, Amplification, Nucleic Acid Electrophoresis, Binding Assay

    Related Articles

    Amplification:

    Article Title: Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform
    Article Snippet: .. Digested amplified products with the restriction enzyme BccI (#R0704S, New England Biolabs) are shown in fig. S2. ..

    Article Title: Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform
    Article Snippet: .. Digested amplified products with the restriction enzyme BccI (#R0704S, New England Biolabs) are shown in . ..

    Article Title: Kinesin Family Member 6 (kif6) Is Necessary for Spine Development in Zebrafish
    Article Snippet: .. Injected embryos and progeny were screened for TALEN-induced mutations by digesting PCR amplified DNA with BccI (New England Biolabs). ..

    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: .. 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.

    Polymerase Chain Reaction:

    Article Title: Pri-Mir-34b/C and Tp-53 Polymorphisms are Associated With The Susceptibility of Papillary Thyroid Carcinoma
    Article Snippet: .. The PCR products were digested with Bcc I (New England BioLabs, Beverly, MA, USA), yielding one band of 147 bp for T allele and two bands of 118 and 29 bp for C allele. .. For the TP-53 Arg72Pro, the primer sequences, PCR conditions and restriction enzymes used were previously established., – To classify the genotypes, digested PCR products were separated on polyacrylamide gels and stained with 1.0 g/L argent nitrate.

    Article Title: Genetic Polymorphism and Expression of CXCR4 in Breast Cancer
    Article Snippet: .. CXCR4 Genotyping PCR products were subjected to restriction digestion by incubating with BccI (New England Biolabs, UK) for 4 h at 37°C. .. The enzymatic restriction products were analyzed by electrophoresis on 10% polyacrylamide gel and detected by a nonradioisotopic technique using silver staining.

    Article Title: Kinesin Family Member 6 (kif6) Is Necessary for Spine Development in Zebrafish
    Article Snippet: .. Injected embryos and progeny were screened for TALEN-induced mutations by digesting PCR amplified DNA with BccI (New England Biolabs). ..

    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: .. 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.

    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: .. 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.

    Injection:

    Article Title: Kinesin Family Member 6 (kif6) Is Necessary for Spine Development in Zebrafish
    Article Snippet: .. Injected embryos and progeny were screened for TALEN-induced mutations by digesting PCR amplified DNA with BccI (New England Biolabs). ..

    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: .. 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.

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    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: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction endonucleases bcci/product/New England Biolabs
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    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

    Design and efficiency of kif6 TALENs. A: Genomic sequence showing left and right TALENs that were designed in the second exon of kif6 near the gw326 mutant allele. The TALEN pair flank a BccI restriction site. B: BccI restriction digest of PCR amplified

    Journal: Developmental dynamics : an official publication of the American Association of Anatomists

    Article Title: Kinesin Family Member 6 (kif6) Is Necessary for Spine Development in Zebrafish

    doi: 10.1002/dvdy.24208

    Figure Lengend Snippet: Design and efficiency of kif6 TALENs. A: Genomic sequence showing left and right TALENs that were designed in the second exon of kif6 near the gw326 mutant allele. The TALEN pair flank a BccI restriction site. B: BccI restriction digest of PCR amplified

    Article Snippet: Injected embryos and progeny were screened for TALEN-induced mutations by digesting PCR amplified DNA with BccI (New England Biolabs).

    Techniques: TALENs, Sequencing, Mutagenesis, Polymerase Chain Reaction, Amplification

    Sequence alignment, designed primers and analytical specificity for the detection of P. falciparum . (A) Illustration of the gene kelch 13 highlighting the location of the LAMP-PfK13 amplicon. (B) Alignment of all human-infective Plasmodium species with P. falciparum as reference sequence. Mismatches in the alignment are displayed as AGTC whereas matched nucleotides are represented as dots. The LAMP-PfK13 primers are illustrated on top of the alignment. (C) . (D) Results showing specific amplification of P. falciparum . The restriction enzyme used for this experiment is BccI (# R0704S, New England BioLabs). The cutting point is illustrated with an arrow and the binding region is orange shadowed in (B). M denotes 100 bp DNA ladder (#10488058, Invitrogen). Acronyms are described in (B). (E) ] used as control for the detection of all Plasmodium species. TTP values are displayed in minutes and labelled.

    Journal: Biosensors & bioelectronics

    Article Title: Quantitative and rapid Plasmodium falciparum malaria diagnosis and artemisinin-resistance detection using a CMOS Lab-on-Chip platform

    doi: 10.1016/j.bios.2019.111678

    Figure Lengend Snippet: Sequence alignment, designed primers and analytical specificity for the detection of P. falciparum . (A) Illustration of the gene kelch 13 highlighting the location of the LAMP-PfK13 amplicon. (B) Alignment of all human-infective Plasmodium species with P. falciparum as reference sequence. Mismatches in the alignment are displayed as AGTC whereas matched nucleotides are represented as dots. The LAMP-PfK13 primers are illustrated on top of the alignment. (C) . (D) Results showing specific amplification of P. falciparum . The restriction enzyme used for this experiment is BccI (# R0704S, New England BioLabs). The cutting point is illustrated with an arrow and the binding region is orange shadowed in (B). M denotes 100 bp DNA ladder (#10488058, Invitrogen). Acronyms are described in (B). (E) ] used as control for the detection of all Plasmodium species. TTP values are displayed in minutes and labelled.

    Article Snippet: Digested amplified products with the restriction enzyme BccI (#R0704S, New England Biolabs) are shown in .

    Techniques: Sequencing, Amplification, Binding Assay

    CXCR4 rs2228014 (C/T) genetic polymorphism. (a) Electrophoretic profile of rs2228014 (C/T). BccI restriction enzyme was used for 4 h at 37°C. Polyacrylamide gel 10% stained with silver nitrate. Lane 1, Ladder DNA fragment marker of 100 bp; Lane 2, PCR product of 236 pb; Lane 3, wild-type homozygous genotype of 133 pb and 103 pb (CC); Lane 4, heterozygous genotype of 236 pb, 133 pb, and 103 pb (CT); Lane 5, blank reaction or negative control (reaction without DNA). (b) Genotype distribution for CXCR4 rs2228014 in breast cancer patients.

    Journal: Analytical cellular pathology (Amsterdam)

    Article Title: Genetic Polymorphism and Expression of CXCR4 in Breast Cancer

    doi: 10.1155/2015/289510

    Figure Lengend Snippet: CXCR4 rs2228014 (C/T) genetic polymorphism. (a) Electrophoretic profile of rs2228014 (C/T). BccI restriction enzyme was used for 4 h at 37°C. Polyacrylamide gel 10% stained with silver nitrate. Lane 1, Ladder DNA fragment marker of 100 bp; Lane 2, PCR product of 236 pb; Lane 3, wild-type homozygous genotype of 133 pb and 103 pb (CC); Lane 4, heterozygous genotype of 236 pb, 133 pb, and 103 pb (CT); Lane 5, blank reaction or negative control (reaction without DNA). (b) Genotype distribution for CXCR4 rs2228014 in breast cancer patients.

    Article Snippet: CXCR4 Genotyping PCR products were subjected to restriction digestion by incubating with BccI (New England Biolabs, UK) for 4 h at 37°C.

    Techniques: Staining, Marker, Polymerase Chain Reaction, Negative Control