r0580  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    BsmBI
    Description:
    BsmBI 1 000 units
    Catalog Number:
    r0580l
    Price:
    302
    Size:
    1 000 units
    Category:
    Restriction Enzymes
    Buy from Supplier


    Structured Review

    New England Biolabs r0580
    BsmBI
    BsmBI 1 000 units
    https://www.bioz.com/result/r0580/product/New England Biolabs
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    r0580 - by Bioz Stars, 2020-08
    99/100 stars

    Images

    Related Articles

    Polymerase Chain Reaction:

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

    Clone Assay:

    Article Title: Joint Universal Modular Plasmids (JUMP): A flexible and comprehensive platform for synthetic biology
    Article Snippet: .. Level 1 and 2 assemblies were performed similarly as in previous Modular Cloning publications: 20 femtomoles of all parts and destination vector were cyclically digested —with BsaI (New England Biolabs Ltd) at 37 °C or BsmBI (NEB) at 42 °C— and ligated with T4 DNA ligase (NEB) at 16 °C. ..

    Plasmid Preparation:

    Article Title: Functional Constraint Profiling of a Viral Protein Reveals Discordance of Evolutionary Conservation and Functionality
    Article Snippet: .. The insert was then digested by BsaI (New England Biolabs, Ipswich, MA), whereas the vector was digested by BsmBI (New England Biolabs). .. Ligation was performed for each of the nine libraries with T4 DNA ligase (Life Technologies, Carlsbad, CA) using the corresponding insert and vector.

    Article Title: Systematic Assembly and Genetic Manipulation of the Mouse Hepatitis Virus A59 Genome
    Article Snippet: .. Incubate at 55°C for 2 h. pSMART-MHV-F: 150 μl plasmid, 10 μl BsmBI, 20 μl NEB Buffer 3, 20 μl water. .. Incubate at 55°C for 2 h. pSMART-MHV-G: 150 μl plasmid, 10 μl SfiI, 2 μl BSA, 20 μl NEB Buffer 2, 18 μl water.

    Article Title: Efficient Genome Editing of a Facultative Thermophile Using Mesophilic spCas9
    Article Snippet: .. Annealed oligos and plasmid pWUR_Cas9nt were digested with BspEI and BsmBI (NEB). .. First, BspEI digestion was performed at 37 °C for 15 min, after which BsmBI was added and the mixture was further incubated at 55 °C for 15 min. After gel purification of the digested products, ligation was performed using NEB T4 ligase, and mixtures were transformed into E. coli TG90.

    Article Title: Joint Universal Modular Plasmids (JUMP): A flexible and comprehensive platform for synthetic biology
    Article Snippet: .. Level 1 and 2 assemblies were performed similarly as in previous Modular Cloning publications: 20 femtomoles of all parts and destination vector were cyclically digested —with BsaI (New England Biolabs Ltd) at 37 °C or BsmBI (NEB) at 42 °C— and ligated with T4 DNA ligase (NEB) at 16 °C. ..

    Construct:

    Article Title: Versatile genetic assembly system (VEGAS) to assemble pathways for expression in S. cerevisiae
    Article Snippet: .. Terminal homology VEGAS ∼1 μg of yGG-assembled, VA-flanked TU constructs were digested with BsmBI (New England Biolabs, R0580) in a final volume of 20 μl. .. 2 μl (∼100 ng) of each digestion product was used directly for yeast transformation along with ∼50 ng of BsaI-linearized VEGAS assembly vector (pJC170 for all experiments described in this work).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs bsmbi
    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 <t>BsaI</t> digestion. The corresponding vector was generated by high-fidelity PCR and <t>BsmBI</t> 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.
    Bsmbi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 219 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsmbi/product/New England Biolabs
    Average 99 stars, based on 219 article reviews
    Price from $9.99 to $1999.99
    bsmbi - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    Image Search Results


    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

    JUMP design and secondary sites. 2A) In SEVA (Standard European Vector Architecture) plasmids, three common short DNA sequences (black) flank three variable regions (coloured). Variable regions are the OriV (origin of replication), AbR (antibiotic selection marker) and “cargo” (any expression cassette). The invariable regions are two transcription terminators flanking the cargo (T1 and T0,) and origin of conjugation (oriT). Invariable regions also contain rare cutting sites, forbidden in the sequence of variable regions. 2B) JUMP is designed as special cargo of SEVA vectors to allow compatibility with future OriV’s and AbR’s of the collection. The cargo contains Upstream Module (outwards AarI); BioBricks prefix (XbaI, EcoRI); Main Module (a screening reporter gene flanked by outwards BsaI and inwards BsmBI for level 1, and vice-versa for level 2); BioBrick suffix (SpeI, PstI); and Downstream Module (outwards BbsI). SEVA’s canonical SpeI site was removed to allow BioBricks compatibility. 2C) Building constructs to test similar genes (G 1 to G n ) as sequences of interest (SOI) that depend on common auxiliary factors (AF) with conventional MoClo might require multiple assembly steps per SOI. D) Introduction of the auxiliary factors in vector chassis using orthogonal use of secondary modules.

    Journal: bioRxiv

    Article Title: Joint Universal Modular Plasmids (JUMP): A flexible and comprehensive platform for synthetic biology

    doi: 10.1101/799585

    Figure Lengend Snippet: JUMP design and secondary sites. 2A) In SEVA (Standard European Vector Architecture) plasmids, three common short DNA sequences (black) flank three variable regions (coloured). Variable regions are the OriV (origin of replication), AbR (antibiotic selection marker) and “cargo” (any expression cassette). The invariable regions are two transcription terminators flanking the cargo (T1 and T0,) and origin of conjugation (oriT). Invariable regions also contain rare cutting sites, forbidden in the sequence of variable regions. 2B) JUMP is designed as special cargo of SEVA vectors to allow compatibility with future OriV’s and AbR’s of the collection. The cargo contains Upstream Module (outwards AarI); BioBricks prefix (XbaI, EcoRI); Main Module (a screening reporter gene flanked by outwards BsaI and inwards BsmBI for level 1, and vice-versa for level 2); BioBrick suffix (SpeI, PstI); and Downstream Module (outwards BbsI). SEVA’s canonical SpeI site was removed to allow BioBricks compatibility. 2C) Building constructs to test similar genes (G 1 to G n ) as sequences of interest (SOI) that depend on common auxiliary factors (AF) with conventional MoClo might require multiple assembly steps per SOI. D) Introduction of the auxiliary factors in vector chassis using orthogonal use of secondary modules.

    Article Snippet: Level 1 and 2 assemblies were performed similarly as in previous Modular Cloning publications: 20 femtomoles of all parts and destination vector were cyclically digested —with BsaI (New England Biolabs Ltd) at 37 °C or BsmBI (NEB) at 42 °C— and ligated with T4 DNA ligase (NEB) at 16 °C.

    Techniques: Plasmid Preparation, Selection, Marker, Expressing, Conjugation Assay, Sequencing, Construct

    Yeast Golden Gate (yGG) to assemble transcription units (TUs) flanked by VEGAS adapters. ( A ) yGG reactions to build TUs destined for VEGAS pathway assembly in S. cerevisiae include five parts: a left VEGAS adapter (LVA), a promoter (PRO), a coding sequence (CDS), a terminator (TER) and a right VEGAS adapter (RVA). Each part is flanked by inwardly facing recognition sequences for the BsaI restriction enzyme, an ‘offset cutter’ which cuts outside its recognition sequence (at positions 1/5 bp downstream) to expose the indicated four base-pair overhangs. All parts are cloned into vectors encoding kanamycin resistance (KAN R ) and an E. coli replication origin (Ori). ( B ) The yGG acceptor vector for VEGAS is designed such that outwardly facing BsaI sites expose overhangs corresponding to the 5′ LVA and 3′ RVA overhangs to promote assembly of the TU in the vector during a one-pot restriction-digestion reaction. The RFP cassette, built for expression in E. coli , is cut out of the vector when a TU correctly assembles, enabling white–red screening. The yGG acceptor vector encodes resistance to ampicillin (AMP R ) ( C ) The structure of a VA-flanked TU assembled by yGG. An assembled TU plus the flanking VA sequences may be released from the yGG acceptor vector by digestion with BsmBI.

    Journal: Nucleic Acids Research

    Article Title: Versatile genetic assembly system (VEGAS) to assemble pathways for expression in S. cerevisiae

    doi: 10.1093/nar/gkv466

    Figure Lengend Snippet: Yeast Golden Gate (yGG) to assemble transcription units (TUs) flanked by VEGAS adapters. ( A ) yGG reactions to build TUs destined for VEGAS pathway assembly in S. cerevisiae include five parts: a left VEGAS adapter (LVA), a promoter (PRO), a coding sequence (CDS), a terminator (TER) and a right VEGAS adapter (RVA). Each part is flanked by inwardly facing recognition sequences for the BsaI restriction enzyme, an ‘offset cutter’ which cuts outside its recognition sequence (at positions 1/5 bp downstream) to expose the indicated four base-pair overhangs. All parts are cloned into vectors encoding kanamycin resistance (KAN R ) and an E. coli replication origin (Ori). ( B ) The yGG acceptor vector for VEGAS is designed such that outwardly facing BsaI sites expose overhangs corresponding to the 5′ LVA and 3′ RVA overhangs to promote assembly of the TU in the vector during a one-pot restriction-digestion reaction. The RFP cassette, built for expression in E. coli , is cut out of the vector when a TU correctly assembles, enabling white–red screening. The yGG acceptor vector encodes resistance to ampicillin (AMP R ) ( C ) The structure of a VA-flanked TU assembled by yGG. An assembled TU plus the flanking VA sequences may be released from the yGG acceptor vector by digestion with BsmBI.

    Article Snippet: Terminal homology VEGAS ∼1 μg of yGG-assembled, VA-flanked TU constructs were digested with BsmBI (New England Biolabs, R0580) in a final volume of 20 μl.

    Techniques: Sequencing, Clone Assay, Plasmid Preparation, Expressing

    VEGAS with adapter homology to assemble the carotenoid pathway in S. cerevisiae . ( A ) The four β-carotene pathway genes ( crtE, crtI, crtYB and tHMG1 ), assembled as TUs flanked by the indicated VAs (see Table 2 for PRO and TER parts), were released from the yGG acceptor vector with BsmBI digestion and co-transformed into yeast with the linearized VEGAS assembly vector. ( B ) S. cerevisiae colonies encoding assembled pathways develop a bright yellow color on medium lacking uracil (SC–Ura; left panel) as well as on YPD medium supplemented with G418 (right panel).

    Journal: Nucleic Acids Research

    Article Title: Versatile genetic assembly system (VEGAS) to assemble pathways for expression in S. cerevisiae

    doi: 10.1093/nar/gkv466

    Figure Lengend Snippet: VEGAS with adapter homology to assemble the carotenoid pathway in S. cerevisiae . ( A ) The four β-carotene pathway genes ( crtE, crtI, crtYB and tHMG1 ), assembled as TUs flanked by the indicated VAs (see Table 2 for PRO and TER parts), were released from the yGG acceptor vector with BsmBI digestion and co-transformed into yeast with the linearized VEGAS assembly vector. ( B ) S. cerevisiae colonies encoding assembled pathways develop a bright yellow color on medium lacking uracil (SC–Ura; left panel) as well as on YPD medium supplemented with G418 (right panel).

    Article Snippet: Terminal homology VEGAS ∼1 μg of yGG-assembled, VA-flanked TU constructs were digested with BsmBI (New England Biolabs, R0580) in a final volume of 20 μl.

    Techniques: Plasmid Preparation, Transformation Assay

    VEGAS with adapter homology to assemble a five-gene pathway. ( A ) The pathway consisting of VA-flanked TUs assembled by yGG may be released in one piece from the yGG acceptor vector by digestion with BsmBI (scissors). ( B ) A genetic pathway may be assembled into the linearized VEGAS assembly vector in S. cerevisiae by homologous recombination between VAs that flank TUs (TU1–5). X's indicate homologous recombination.

    Journal: Nucleic Acids Research

    Article Title: Versatile genetic assembly system (VEGAS) to assemble pathways for expression in S. cerevisiae

    doi: 10.1093/nar/gkv466

    Figure Lengend Snippet: VEGAS with adapter homology to assemble a five-gene pathway. ( A ) The pathway consisting of VA-flanked TUs assembled by yGG may be released in one piece from the yGG acceptor vector by digestion with BsmBI (scissors). ( B ) A genetic pathway may be assembled into the linearized VEGAS assembly vector in S. cerevisiae by homologous recombination between VAs that flank TUs (TU1–5). X's indicate homologous recombination.

    Article Snippet: Terminal homology VEGAS ∼1 μg of yGG-assembled, VA-flanked TU constructs were digested with BsmBI (New England Biolabs, R0580) in a final volume of 20 μl.

    Techniques: Plasmid Preparation, Homologous Recombination

    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