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  • 99
    New England Biolabs bsmb1 restriction enzyme
    Schematic diagram of the RNA interference expression vector pRNAi-U6H1/Neo. The vector contains a multiple cloning sites with two <t>BsmB</t> I restriction sites to permit cloning of <t>Fok</t> I-digested sRNA fragments and an Sfi I site to assist in screening for recombinants.
    Bsmb1 Restriction Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 103 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    New England Biolabs esp3i bsmbi cut ligations
    Schematic diagram of the RNA interference expression vector pRNAi-U6H1/Neo. The vector contains a multiple cloning sites with two <t>BsmB</t> I restriction sites to permit cloning of <t>Fok</t> I-digested sRNA fragments and an Sfi I site to assist in screening for recombinants.
    Esp3i Bsmbi Cut Ligations, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs type iis restriction enzymes
    Comparison of type II and <t>IIS</t> restriction enzyme-mediated protein tagging strategy ( a ) Type II restriction enzymes (TII-es) recognize palindromic DNA sequences. For example, <t>EcoRI</t> recognizes 5′-GAATTC-3′ (marked by top curly bracket) and creates 4 base pairs overhangs highlighted in red. ( b ) Single or double type II restriction enzymes cassette (highlighted in blue box) for traditional protein tagging. Note that in all destination clones, varying junction sequences exist adjacent to both sides of the tag. ( c ) Type IIS restriction enzymes (TIIS-es) recognize non-palindromic, asymmetrical DNA sequences. For example, BsaI recognizes 5′-GGTCTC-3′ (marked by top curly bracket) and cleaves DNA one bp away (indicated by the two arrows), producing N 2 ′N 3 ′N 4 ′N 5 ′ custom sticky end (highlighted in red). N indicates four bases of DNA, including A, T, G and C. Apostrophe (’) indicates the complementary base of the DNA. ( d ) Type IIS restriction enzyme DNA cassette (TIIS DNA cassette highlighted in blue box) for precision tagging. Note that on both ends of a tag, the flanking sequences (such as BsaI-released 5′-N 2 ′N 3 ′N 4 ′N 5 ′ and 5′-N 2 N 3 N 4 N 5 belong to gene-specific sequences including SP or gene of interest indicated by two closely dotted lines. After Tag replaces type IIS DNA cassette, a scarless tagging clone can be generated. Comparison between traditional and precision tagging were summarized in the bottom table. * Gibson assembly sometimes fails due to certain DNA sequences such as repetitive region or creating one or two nucleotides deletion.
    Type Iis Restriction Enzymes, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
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    99
    New England Biolabs restriction endonuclease bsmbi
    Comparison of type II and <t>IIS</t> restriction enzyme-mediated protein tagging strategy ( a ) Type II restriction enzymes (TII-es) recognize palindromic DNA sequences. For example, <t>EcoRI</t> recognizes 5′-GAATTC-3′ (marked by top curly bracket) and creates 4 base pairs overhangs highlighted in red. ( b ) Single or double type II restriction enzymes cassette (highlighted in blue box) for traditional protein tagging. Note that in all destination clones, varying junction sequences exist adjacent to both sides of the tag. ( c ) Type IIS restriction enzymes (TIIS-es) recognize non-palindromic, asymmetrical DNA sequences. For example, BsaI recognizes 5′-GGTCTC-3′ (marked by top curly bracket) and cleaves DNA one bp away (indicated by the two arrows), producing N 2 ′N 3 ′N 4 ′N 5 ′ custom sticky end (highlighted in red). N indicates four bases of DNA, including A, T, G and C. Apostrophe (’) indicates the complementary base of the DNA. ( d ) Type IIS restriction enzyme DNA cassette (TIIS DNA cassette highlighted in blue box) for precision tagging. Note that on both ends of a tag, the flanking sequences (such as BsaI-released 5′-N 2 ′N 3 ′N 4 ′N 5 ′ and 5′-N 2 N 3 N 4 N 5 belong to gene-specific sequences including SP or gene of interest indicated by two closely dotted lines. After Tag replaces type IIS DNA cassette, a scarless tagging clone can be generated. Comparison between traditional and precision tagging were summarized in the bottom table. * Gibson assembly sometimes fails due to certain DNA sequences such as repetitive region or creating one or two nucleotides deletion.
    Restriction Endonuclease Bsmbi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs dna modification enzymes
    Comparison of type II and <t>IIS</t> restriction enzyme-mediated protein tagging strategy ( a ) Type II restriction enzymes (TII-es) recognize palindromic DNA sequences. For example, <t>EcoRI</t> recognizes 5′-GAATTC-3′ (marked by top curly bracket) and creates 4 base pairs overhangs highlighted in red. ( b ) Single or double type II restriction enzymes cassette (highlighted in blue box) for traditional protein tagging. Note that in all destination clones, varying junction sequences exist adjacent to both sides of the tag. ( c ) Type IIS restriction enzymes (TIIS-es) recognize non-palindromic, asymmetrical DNA sequences. For example, BsaI recognizes 5′-GGTCTC-3′ (marked by top curly bracket) and cleaves DNA one bp away (indicated by the two arrows), producing N 2 ′N 3 ′N 4 ′N 5 ′ custom sticky end (highlighted in red). N indicates four bases of DNA, including A, T, G and C. Apostrophe (’) indicates the complementary base of the DNA. ( d ) Type IIS restriction enzyme DNA cassette (TIIS DNA cassette highlighted in blue box) for precision tagging. Note that on both ends of a tag, the flanking sequences (such as BsaI-released 5′-N 2 ′N 3 ′N 4 ′N 5 ′ and 5′-N 2 N 3 N 4 N 5 belong to gene-specific sequences including SP or gene of interest indicated by two closely dotted lines. After Tag replaces type IIS DNA cassette, a scarless tagging clone can be generated. Comparison between traditional and precision tagging were summarized in the bottom table. * Gibson assembly sometimes fails due to certain DNA sequences such as repetitive region or creating one or two nucleotides deletion.
    Dna Modification Enzymes, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 562 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 562 article reviews
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    Image Search Results


    Schematic diagram of the RNA interference expression vector pRNAi-U6H1/Neo. The vector contains a multiple cloning sites with two BsmB I restriction sites to permit cloning of Fok I-digested sRNA fragments and an Sfi I site to assist in screening for recombinants.

    Journal: Nucleic Acid Therapeutics

    Article Title: Size Unbiased Representative Enzymatically Generated RNAi (SURER) Library and Application for RNAi Therapeutic Screens

    doi: 10.1089/nat.2014.0514

    Figure Lengend Snippet: Schematic diagram of the RNA interference expression vector pRNAi-U6H1/Neo. The vector contains a multiple cloning sites with two BsmB I restriction sites to permit cloning of Fok I-digested sRNA fragments and an Sfi I site to assist in screening for recombinants.

    Article Snippet: Fok I-digested fragments were ligated into linearized pRNAi-U6H1/Neo. pRNAi-U6H1/Neo was digested with BsmB I and dephosphorylated with calf intestinal alkaline phosphatase according to manufacturer's (NEB) protocols.

    Techniques: Expressing, Plasmid Preparation, Clone Assay

    Construction of the mutant libraries. (A) A schematic representation of the fitness profiling experiment is shown. A 240 bp insert was generated by error-prone PCR and BsaI digestion. The corresponding vector was generated by high-fidelity PCR and BsmBI digestion. Each of the nine plasmid libraries in this study consist of ∼ 50,000 clones. Each viral mutant library was rescued by transfecting ∼ 35 million 293T cells. Each infection was performed with ∼ 10 million A549 cells. (B) A schematic representation of the sequencing library preparation is shown. DNA plasmid mutant library or viral cDNA was used for PCR. This PCR amplified the 240 bp randomized region. The amplicon product was then digested with BpmI, end-repaired, dA-tailed, ligated to sequencing adapters, and sequenced using the Illumina MiSeq platform. BpmI digestion removed the primer region in the amplicon PCR, resulting in sequencing reads covering only the barcode for multiplex sequencing and the 240 bp region that was randomized in the mutant library. With this experimental design, the number of mutations carried by individual genomes in the mutant libraries could be precisely determined.

    Journal: PLoS Genetics

    Article Title: Functional Constraint Profiling of a Viral Protein Reveals Discordance of Evolutionary Conservation and Functionality

    doi: 10.1371/journal.pgen.1005310

    Figure Lengend Snippet: Construction of the mutant libraries. (A) A schematic representation of the fitness profiling experiment is shown. A 240 bp insert was generated by error-prone PCR and BsaI digestion. The corresponding vector was generated by high-fidelity PCR and BsmBI digestion. Each of the nine plasmid libraries in this study consist of ∼ 50,000 clones. Each viral mutant library was rescued by transfecting ∼ 35 million 293T cells. Each infection was performed with ∼ 10 million A549 cells. (B) A schematic representation of the sequencing library preparation is shown. DNA plasmid mutant library or viral cDNA was used for PCR. This PCR amplified the 240 bp randomized region. The amplicon product was then digested with BpmI, end-repaired, dA-tailed, ligated to sequencing adapters, and sequenced using the Illumina MiSeq platform. BpmI digestion removed the primer region in the amplicon PCR, resulting in sequencing reads covering only the barcode for multiplex sequencing and the 240 bp region that was randomized in the mutant library. With this experimental design, the number of mutations carried by individual genomes in the mutant libraries could be precisely determined.

    Article Snippet: The insert was then digested by BsaI (New England Biolabs, Ipswich, MA), whereas the vector was digested by BsmBI (New England Biolabs).

    Techniques: Mutagenesis, Generated, Polymerase Chain Reaction, Plasmid Preparation, Clone Assay, Infection, Sequencing, Amplification, Multiplex Assay

    gRNA library construction using a semi-random primer. ( A ) Semi-random primer. Poly(A), polyadenylate. ( B ) Type III and type IIS restriction sites to cut out the 20-bp guide sequence. Ec, Eco P15I; Ac, Acu I. ( C ) Scheme of gRNA library construction. Bg, Bgl II; Xb, Xba I; Bs, Bsm BI; Aa, Aat II. PCR, polymerase chain reaction; lentiCRISPR v2, lentiCRISPR version 2. ( D ) Short-range PCR for PCR cycle optimization and size fractionation of the guide sequence. PCR products were run on 20% polyacrylamide gels. A 10-bp ladder was used as the size marker. Bands of the expected sizes are marked by triangles.

    Journal: Science Advances

    Article Title: A method to convert mRNA into a gRNA library for CRISPR/Cas9 editing of any organism

    doi: 10.1126/sciadv.1600699

    Figure Lengend Snippet: gRNA library construction using a semi-random primer. ( A ) Semi-random primer. Poly(A), polyadenylate. ( B ) Type III and type IIS restriction sites to cut out the 20-bp guide sequence. Ec, Eco P15I; Ac, Acu I. ( C ) Scheme of gRNA library construction. Bg, Bgl II; Xb, Xba I; Bs, Bsm BI; Aa, Aat II. PCR, polymerase chain reaction; lentiCRISPR v2, lentiCRISPR version 2. ( D ) Short-range PCR for PCR cycle optimization and size fractionation of the guide sequence. PCR products were run on 20% polyacrylamide gels. A 10-bp ladder was used as the size marker. Bands of the expected sizes are marked by triangles.

    Article Snippet: Bsm BI/Aat II digestion The PCR product was digested with 10 μl of Bsm BI (10 U/μl; NEB) in 1× NEBuffer 3.1 in a 100-μl volume at 55°C for 6 hours, and then 5 μl of Aat II (20 U/μl; NEB) was added to the solution, which was left at 37°C overnight.

    Techniques: Sequencing, Polymerase Chain Reaction, Fractionation, Marker

    Comparison of type II and IIS restriction enzyme-mediated protein tagging strategy ( a ) Type II restriction enzymes (TII-es) recognize palindromic DNA sequences. For example, EcoRI recognizes 5′-GAATTC-3′ (marked by top curly bracket) and creates 4 base pairs overhangs highlighted in red. ( b ) Single or double type II restriction enzymes cassette (highlighted in blue box) for traditional protein tagging. Note that in all destination clones, varying junction sequences exist adjacent to both sides of the tag. ( c ) Type IIS restriction enzymes (TIIS-es) recognize non-palindromic, asymmetrical DNA sequences. For example, BsaI recognizes 5′-GGTCTC-3′ (marked by top curly bracket) and cleaves DNA one bp away (indicated by the two arrows), producing N 2 ′N 3 ′N 4 ′N 5 ′ custom sticky end (highlighted in red). N indicates four bases of DNA, including A, T, G and C. Apostrophe (’) indicates the complementary base of the DNA. ( d ) Type IIS restriction enzyme DNA cassette (TIIS DNA cassette highlighted in blue box) for precision tagging. Note that on both ends of a tag, the flanking sequences (such as BsaI-released 5′-N 2 ′N 3 ′N 4 ′N 5 ′ and 5′-N 2 N 3 N 4 N 5 belong to gene-specific sequences including SP or gene of interest indicated by two closely dotted lines. After Tag replaces type IIS DNA cassette, a scarless tagging clone can be generated. Comparison between traditional and precision tagging were summarized in the bottom table. * Gibson assembly sometimes fails due to certain DNA sequences such as repetitive region or creating one or two nucleotides deletion.

    Journal: Biochemical and biophysical research communications

    Article Title: Highly Efficient One-Step Scarless Protein Tagging by Type IIS Restriction Endonuclease-Mediated Precision Cloning

    doi: 10.1016/j.bbrc.2017.05.153

    Figure Lengend Snippet: Comparison of type II and IIS restriction enzyme-mediated protein tagging strategy ( a ) Type II restriction enzymes (TII-es) recognize palindromic DNA sequences. For example, EcoRI recognizes 5′-GAATTC-3′ (marked by top curly bracket) and creates 4 base pairs overhangs highlighted in red. ( b ) Single or double type II restriction enzymes cassette (highlighted in blue box) for traditional protein tagging. Note that in all destination clones, varying junction sequences exist adjacent to both sides of the tag. ( c ) Type IIS restriction enzymes (TIIS-es) recognize non-palindromic, asymmetrical DNA sequences. For example, BsaI recognizes 5′-GGTCTC-3′ (marked by top curly bracket) and cleaves DNA one bp away (indicated by the two arrows), producing N 2 ′N 3 ′N 4 ′N 5 ′ custom sticky end (highlighted in red). N indicates four bases of DNA, including A, T, G and C. Apostrophe (’) indicates the complementary base of the DNA. ( d ) Type IIS restriction enzyme DNA cassette (TIIS DNA cassette highlighted in blue box) for precision tagging. Note that on both ends of a tag, the flanking sequences (such as BsaI-released 5′-N 2 ′N 3 ′N 4 ′N 5 ′ and 5′-N 2 N 3 N 4 N 5 belong to gene-specific sequences including SP or gene of interest indicated by two closely dotted lines. After Tag replaces type IIS DNA cassette, a scarless tagging clone can be generated. Comparison between traditional and precision tagging were summarized in the bottom table. * Gibson assembly sometimes fails due to certain DNA sequences such as repetitive region or creating one or two nucleotides deletion.

    Article Snippet: All the enzymes including type II restriction enzymes (EcoRI, BamHI, SalI and Bau36I), type IIS restriction enzymes (BsaI, BbsI and BsmBI), T4 DNA ligase, Phusion enzyme, and T5 exonuclease were purchased from New England BioLabs.

    Techniques: Clone Assay, Generated