biobrick assembly kit  (New England Biolabs)


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    Structured Review

    New England Biolabs biobrick assembly kit
    Examples of <t>biobrick</t> assemblies. (A) Structure of a classical plasmid for EGFP expression using 5 biobricks. (B) Structure of bicistronic custom plasmid. 8 biobricks are linked together to make a construct for C-terminal fusions to the blue fluorescent protein, cerulean, using classical restriction enzyme multiple cloning sites (MCS). For illustration, the actin-bundling binding protein sEspin is cloned into the MCS, resulting in a fusion with cerulean. (C) Fluorescence microscopy image of the EGFP construct in panel A, after transient transfection into HEK293 cells. (D) A fluorescence microscopy image of the Cerulean-sEspin fusion construct in panel B, allows the visualisation of stress fiber-like structures in a HEK293T cell. Scale bars are indicated below each image.
    Biobrick Assembly Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biobrick assembly kit/product/New England Biolabs
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    biobrick assembly kit - by Bioz Stars, 2022-09
    93/100 stars

    Images

    1) Product Images from "A Biobrick Library for Cloning Custom Eukaryotic Plasmids"

    Article Title: A Biobrick Library for Cloning Custom Eukaryotic Plasmids

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0023685

    Examples of biobrick assemblies. (A) Structure of a classical plasmid for EGFP expression using 5 biobricks. (B) Structure of bicistronic custom plasmid. 8 biobricks are linked together to make a construct for C-terminal fusions to the blue fluorescent protein, cerulean, using classical restriction enzyme multiple cloning sites (MCS). For illustration, the actin-bundling binding protein sEspin is cloned into the MCS, resulting in a fusion with cerulean. (C) Fluorescence microscopy image of the EGFP construct in panel A, after transient transfection into HEK293 cells. (D) A fluorescence microscopy image of the Cerulean-sEspin fusion construct in panel B, allows the visualisation of stress fiber-like structures in a HEK293T cell. Scale bars are indicated below each image.
    Figure Legend Snippet: Examples of biobrick assemblies. (A) Structure of a classical plasmid for EGFP expression using 5 biobricks. (B) Structure of bicistronic custom plasmid. 8 biobricks are linked together to make a construct for C-terminal fusions to the blue fluorescent protein, cerulean, using classical restriction enzyme multiple cloning sites (MCS). For illustration, the actin-bundling binding protein sEspin is cloned into the MCS, resulting in a fusion with cerulean. (C) Fluorescence microscopy image of the EGFP construct in panel A, after transient transfection into HEK293 cells. (D) A fluorescence microscopy image of the Cerulean-sEspin fusion construct in panel B, allows the visualisation of stress fiber-like structures in a HEK293T cell. Scale bars are indicated below each image.

    Techniques Used: Plasmid Preparation, Expressing, Construct, Clone Assay, Binding Assay, Fluorescence, Microscopy, Transfection

    The biobrick assembly principle [8] , [10] . (A) Each biobrick part has the same prefix and suffix, containing restriction enzyme sites. (B) Following restriction digests, a two-insert ligation into the biobrick vector results in a biobrick fusion. (C) The new biobrick part regenerates the original prefix and suffix, but contains an in-frame Thr-Arg scar in protein-coding fusions. (D) MS2 binding site concatemers (MS2 BS), built with iterative biobrick assembly, from 1 to 12-copies (4 steps). M = marker (1 kb ladder). The upstream and downstream sequences between the primer annealing sites and the biobricks contribute 312 bp, while each MS2 BS is 39 bp.
    Figure Legend Snippet: The biobrick assembly principle [8] , [10] . (A) Each biobrick part has the same prefix and suffix, containing restriction enzyme sites. (B) Following restriction digests, a two-insert ligation into the biobrick vector results in a biobrick fusion. (C) The new biobrick part regenerates the original prefix and suffix, but contains an in-frame Thr-Arg scar in protein-coding fusions. (D) MS2 binding site concatemers (MS2 BS), built with iterative biobrick assembly, from 1 to 12-copies (4 steps). M = marker (1 kb ladder). The upstream and downstream sequences between the primer annealing sites and the biobricks contribute 312 bp, while each MS2 BS is 39 bp.

    Techniques Used: Ligation, Plasmid Preparation, Binding Assay, Marker

    Multiple Cloning Site (MCS) biobricks. The uniqueness of each cloning site is dependent on whether the remaining biobricks and backbones used for the custom plasmid also contain the restriction site. Blunt end restriction enzymes are represented in italic. These biobricks link classical cloning to the biobrick system.
    Figure Legend Snippet: Multiple Cloning Site (MCS) biobricks. The uniqueness of each cloning site is dependent on whether the remaining biobricks and backbones used for the custom plasmid also contain the restriction site. Blunt end restriction enzymes are represented in italic. These biobricks link classical cloning to the biobrick system.

    Techniques Used: Clone Assay, Plasmid Preparation

    2) Product Images from "Introduction of customized inserts for streamlined assembly and optimization of BioBrick synthetic genetic circuits"

    Article Title: Introduction of customized inserts for streamlined assembly and optimization of BioBrick synthetic genetic circuits

    Journal: Journal of Biological Engineering

    doi: 10.1186/1754-1611-4-17

    Desired BioBrick circuit modifications and approach with BioScaffolds . Every BioBrick standard biological part (a) consists of a DNA sequence embedded between a
    Figure Legend Snippet: Desired BioBrick circuit modifications and approach with BioScaffolds . Every BioBrick standard biological part (a) consists of a DNA sequence embedded between a "prefix" sequence (purple box) and a "suffix" sequence (yellow box). Parts may also contain scars (b) , which form when two parts, such as "A1" and "A2" in (c) are fused together using BioBrick assembly [ 12 ]. In many cases one would like to convert an undesired scar between two parts in a BioBrick assembly into a different part or completely remove it (d) . Our approach is to create a new BioBrick part (the BioScaffold) (e) . The BioScaffold can be assembled into a circuit using BioBrick assembly, but unlike normal BioBricks it can be removed and replaced with a new part (f) . In this paper we develop a single prototype BioScaffold that illustrates how BioScaffolds can be used to either insert parts or remove scars.

    Techniques Used: Sequencing

    3) Product Images from "Biotechnology by Design: An Introductory Level, Project-Based, Synthetic Biology Laboratory Program for Undergraduate Students †"

    Article Title: Biotechnology by Design: An Introductory Level, Project-Based, Synthetic Biology Laboratory Program for Undergraduate Students †

    Journal: Journal of Microbiology & Biology Education

    doi: 10.1128/jmbe.v16i2.971

    BioBrick Assembly Methods Flowchart. The flowchart illustrates the stepwise construction of the reporter plasmid using a Back-End Assembly approach. Molecular biology techniques are listed in blue type, and decision points are outlined in red. Verification
    Figure Legend Snippet: BioBrick Assembly Methods Flowchart. The flowchart illustrates the stepwise construction of the reporter plasmid using a Back-End Assembly approach. Molecular biology techniques are listed in blue type, and decision points are outlined in red. Verification

    Techniques Used: Plasmid Preparation

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    New England Biolabs biobrick assembly kit
    Examples of <t>biobrick</t> assemblies. (A) Structure of a classical plasmid for EGFP expression using 5 biobricks. (B) Structure of bicistronic custom plasmid. 8 biobricks are linked together to make a construct for C-terminal fusions to the blue fluorescent protein, cerulean, using classical restriction enzyme multiple cloning sites (MCS). For illustration, the actin-bundling binding protein sEspin is cloned into the MCS, resulting in a fusion with cerulean. (C) Fluorescence microscopy image of the EGFP construct in panel A, after transient transfection into HEK293 cells. (D) A fluorescence microscopy image of the Cerulean-sEspin fusion construct in panel B, allows the visualisation of stress fiber-like structures in a HEK293T cell. Scale bars are indicated below each image.
    Biobrick Assembly Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biobrick assembly kit/product/New England Biolabs
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    biobrick assembly kit - by Bioz Stars, 2022-09
    93/100 stars
      Buy from Supplier

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    Examples of biobrick assemblies. (A) Structure of a classical plasmid for EGFP expression using 5 biobricks. (B) Structure of bicistronic custom plasmid. 8 biobricks are linked together to make a construct for C-terminal fusions to the blue fluorescent protein, cerulean, using classical restriction enzyme multiple cloning sites (MCS). For illustration, the actin-bundling binding protein sEspin is cloned into the MCS, resulting in a fusion with cerulean. (C) Fluorescence microscopy image of the EGFP construct in panel A, after transient transfection into HEK293 cells. (D) A fluorescence microscopy image of the Cerulean-sEspin fusion construct in panel B, allows the visualisation of stress fiber-like structures in a HEK293T cell. Scale bars are indicated below each image.

    Journal: PLoS ONE

    Article Title: A Biobrick Library for Cloning Custom Eukaryotic Plasmids

    doi: 10.1371/journal.pone.0023685

    Figure Lengend Snippet: Examples of biobrick assemblies. (A) Structure of a classical plasmid for EGFP expression using 5 biobricks. (B) Structure of bicistronic custom plasmid. 8 biobricks are linked together to make a construct for C-terminal fusions to the blue fluorescent protein, cerulean, using classical restriction enzyme multiple cloning sites (MCS). For illustration, the actin-bundling binding protein sEspin is cloned into the MCS, resulting in a fusion with cerulean. (C) Fluorescence microscopy image of the EGFP construct in panel A, after transient transfection into HEK293 cells. (D) A fluorescence microscopy image of the Cerulean-sEspin fusion construct in panel B, allows the visualisation of stress fiber-like structures in a HEK293T cell. Scale bars are indicated below each image.

    Article Snippet: Biobrick assemblies Assemblies were performed similarly to the method presented in the Biobrick assembly kit from New England Biolabs (NEB; Ref. E0546S) and the Gingko Bioworks manual ( http://ginkgobioworks.com/support/ ).

    Techniques: Plasmid Preparation, Expressing, Construct, Clone Assay, Binding Assay, Fluorescence, Microscopy, Transfection

    The biobrick assembly principle [8] , [10] . (A) Each biobrick part has the same prefix and suffix, containing restriction enzyme sites. (B) Following restriction digests, a two-insert ligation into the biobrick vector results in a biobrick fusion. (C) The new biobrick part regenerates the original prefix and suffix, but contains an in-frame Thr-Arg scar in protein-coding fusions. (D) MS2 binding site concatemers (MS2 BS), built with iterative biobrick assembly, from 1 to 12-copies (4 steps). M = marker (1 kb ladder). The upstream and downstream sequences between the primer annealing sites and the biobricks contribute 312 bp, while each MS2 BS is 39 bp.

    Journal: PLoS ONE

    Article Title: A Biobrick Library for Cloning Custom Eukaryotic Plasmids

    doi: 10.1371/journal.pone.0023685

    Figure Lengend Snippet: The biobrick assembly principle [8] , [10] . (A) Each biobrick part has the same prefix and suffix, containing restriction enzyme sites. (B) Following restriction digests, a two-insert ligation into the biobrick vector results in a biobrick fusion. (C) The new biobrick part regenerates the original prefix and suffix, but contains an in-frame Thr-Arg scar in protein-coding fusions. (D) MS2 binding site concatemers (MS2 BS), built with iterative biobrick assembly, from 1 to 12-copies (4 steps). M = marker (1 kb ladder). The upstream and downstream sequences between the primer annealing sites and the biobricks contribute 312 bp, while each MS2 BS is 39 bp.

    Article Snippet: Biobrick assemblies Assemblies were performed similarly to the method presented in the Biobrick assembly kit from New England Biolabs (NEB; Ref. E0546S) and the Gingko Bioworks manual ( http://ginkgobioworks.com/support/ ).

    Techniques: Ligation, Plasmid Preparation, Binding Assay, Marker

    Multiple Cloning Site (MCS) biobricks. The uniqueness of each cloning site is dependent on whether the remaining biobricks and backbones used for the custom plasmid also contain the restriction site. Blunt end restriction enzymes are represented in italic. These biobricks link classical cloning to the biobrick system.

    Journal: PLoS ONE

    Article Title: A Biobrick Library for Cloning Custom Eukaryotic Plasmids

    doi: 10.1371/journal.pone.0023685

    Figure Lengend Snippet: Multiple Cloning Site (MCS) biobricks. The uniqueness of each cloning site is dependent on whether the remaining biobricks and backbones used for the custom plasmid also contain the restriction site. Blunt end restriction enzymes are represented in italic. These biobricks link classical cloning to the biobrick system.

    Article Snippet: Biobrick assemblies Assemblies were performed similarly to the method presented in the Biobrick assembly kit from New England Biolabs (NEB; Ref. E0546S) and the Gingko Bioworks manual ( http://ginkgobioworks.com/support/ ).

    Techniques: Clone Assay, Plasmid Preparation

    Desired BioBrick circuit modifications and approach with BioScaffolds . Every BioBrick standard biological part (a) consists of a DNA sequence embedded between a

    Journal: Journal of Biological Engineering

    Article Title: Introduction of customized inserts for streamlined assembly and optimization of BioBrick synthetic genetic circuits

    doi: 10.1186/1754-1611-4-17

    Figure Lengend Snippet: Desired BioBrick circuit modifications and approach with BioScaffolds . Every BioBrick standard biological part (a) consists of a DNA sequence embedded between a "prefix" sequence (purple box) and a "suffix" sequence (yellow box). Parts may also contain scars (b) , which form when two parts, such as "A1" and "A2" in (c) are fused together using BioBrick assembly [ 12 ]. In many cases one would like to convert an undesired scar between two parts in a BioBrick assembly into a different part or completely remove it (d) . Our approach is to create a new BioBrick part (the BioScaffold) (e) . The BioScaffold can be assembled into a circuit using BioBrick assembly, but unlike normal BioBricks it can be removed and replaced with a new part (f) . In this paper we develop a single prototype BioScaffold that illustrates how BioScaffolds can be used to either insert parts or remove scars.

    Article Snippet: The assembly was performed in two rounds using the BioBrick Assembly kit (New England Biolabs, Inc.) for three antibiotic (3A) assembly.

    Techniques: Sequencing

    BioBrick Assembly Methods Flowchart. The flowchart illustrates the stepwise construction of the reporter plasmid using a Back-End Assembly approach. Molecular biology techniques are listed in blue type, and decision points are outlined in red. Verification

    Journal: Journal of Microbiology & Biology Education

    Article Title: Biotechnology by Design: An Introductory Level, Project-Based, Synthetic Biology Laboratory Program for Undergraduate Students †

    doi: 10.1128/jmbe.v16i2.971

    Figure Lengend Snippet: BioBrick Assembly Methods Flowchart. The flowchart illustrates the stepwise construction of the reporter plasmid using a Back-End Assembly approach. Molecular biology techniques are listed in blue type, and decision points are outlined in red. Verification

    Article Snippet: Three commercial kits are recommended for the required enzymes and DNA ligase (NEB BioBrick Assembly Kit, NEB-E0546), DNA fragment purification (IBI Scientific Gel/PCR DNA Fragment Extraction Kit, IB470300), and plasmid extraction from E. coli cells (IBI Scientific Hi-Speed Mini Plasmid Kit, IB47101).

    Techniques: Plasmid Preparation