gibson assembly master mix  (New England Biolabs)


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  • 99
    Name:
    Gibson Assembly Master Mix
    Description:
    Gibson Assembly Master Mix 50 rxns
    Catalog Number:
    e2611l
    Price:
    649
    Size:
    50 rxns
    Category:
    Cloning and Expression Systems
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    Structured Review

    New England Biolabs gibson assembly master mix
    Gibson Assembly Master Mix
    Gibson Assembly Master Mix 50 rxns
    https://www.bioz.com/result/gibson assembly master mix/product/New England Biolabs
    Average 99 stars, based on 410 article reviews
    Price from $9.99 to $1999.99
    gibson assembly master mix - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "A mixed culture of bacterial cells enables an economic DNA storage on a large scale"

    Article Title: A mixed culture of bacterial cells enables an economic DNA storage on a large scale

    Journal: Communications Biology

    doi: 10.1038/s42003-020-01141-7

    Large-scale DNA data storage in living cells. a The workflow for the manufacture of a mixed culture living cell data storage material. The assembled oligo pool with 10 6 to 10 7 average copies for each oligo was subjected to assembly and then introduced into E. coli cell. A 10 1 to 10 2 average colony number for each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieval and information decoding. b The 0.9% lost oligos in the 1 st passage of the one-fragment assembly (red line) and the 0.56% lost oligos in the 10× deep sequencing reads of the original master pool (blue line) were mapped to the oligo frequency distribution of the original master pool (gray line). c In contrast with previous reported major systems for DNA storage in living cells, including 0.25 kbps by Yachie in 2007, 14.56 bps by Shipman in 2017 and 2.448 kbps by Sun in 2019, the total of 97.728 kbps of DNA for the 509 oligos pool and 2304 Kbps for the 11520 oligos pool stored in a mixed culture of E. coli cells at a cost lower than 0.001$ per base, and the mixed cell storage material could be manufactured within 24 h.
    Figure Legend Snippet: Large-scale DNA data storage in living cells. a The workflow for the manufacture of a mixed culture living cell data storage material. The assembled oligo pool with 10 6 to 10 7 average copies for each oligo was subjected to assembly and then introduced into E. coli cell. A 10 1 to 10 2 average colony number for each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieval and information decoding. b The 0.9% lost oligos in the 1 st passage of the one-fragment assembly (red line) and the 0.56% lost oligos in the 10× deep sequencing reads of the original master pool (blue line) were mapped to the oligo frequency distribution of the original master pool (gray line). c In contrast with previous reported major systems for DNA storage in living cells, including 0.25 kbps by Yachie in 2007, 14.56 bps by Shipman in 2017 and 2.448 kbps by Sun in 2019, the total of 97.728 kbps of DNA for the 509 oligos pool and 2304 Kbps for the 11520 oligos pool stored in a mixed culture of E. coli cells at a cost lower than 0.001$ per base, and the mixed cell storage material could be manufactured within 24 h.

    Techniques Used: Amplification, Plasmid Preparation, Sequencing

    2) Product Images from "Mixed Culture of Bacterial Cell for Large Scale DNA Storage"

    Article Title: Mixed Culture of Bacterial Cell for Large Scale DNA Storage

    Journal: bioRxiv

    doi: 10.1101/2020.02.21.960476

    A large-scale DNA data storage in living cell. a) The workflow for the manufacture of mixed culture living cell data storage materials. Oligo pool was assembled with 1E+6⁓7 of average copy of each oligo was subjected to assembly and then transformed into E. coli cell with about 1E+1⁓2 average colony number of each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieve and information decoding. b) the 0.9% dropout oligos in 1 st passaging of one fragment assembly (red line) and the 0.56% dropout oligos in 10x sequencing reads of original master pool (blue line) were mapped to the oligo frequency distribution of original master pool (gray line). c) In comparison with previous reported major systems for DNA storage in living cell including 0.25 kbps by Yachie in 2007, 18.2 bps by Shipman in 2017 and 2.8 kbps by Sun in 2019, totally 97.7 kbps DNA for 509 oligos pool and 2304 kbps for 11520 oligos pool were stored in mixed culture of E. coli cells at cost lower than 1E-4$ per base and mixed cell storage materials could be manufactured within 24 hrs.
    Figure Legend Snippet: A large-scale DNA data storage in living cell. a) The workflow for the manufacture of mixed culture living cell data storage materials. Oligo pool was assembled with 1E+6⁓7 of average copy of each oligo was subjected to assembly and then transformed into E. coli cell with about 1E+1⁓2 average colony number of each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieve and information decoding. b) the 0.9% dropout oligos in 1 st passaging of one fragment assembly (red line) and the 0.56% dropout oligos in 10x sequencing reads of original master pool (blue line) were mapped to the oligo frequency distribution of original master pool (gray line). c) In comparison with previous reported major systems for DNA storage in living cell including 0.25 kbps by Yachie in 2007, 18.2 bps by Shipman in 2017 and 2.8 kbps by Sun in 2019, totally 97.7 kbps DNA for 509 oligos pool and 2304 kbps for 11520 oligos pool were stored in mixed culture of E. coli cells at cost lower than 1E-4$ per base and mixed cell storage materials could be manufactured within 24 hrs.

    Techniques Used: Transformation Assay, Amplification, Plasmid Preparation, Passaging, Sequencing

    3) Product Images from "A mixed culture of bacterial cells enables an economic DNA storage on a large scale"

    Article Title: A mixed culture of bacterial cells enables an economic DNA storage on a large scale

    Journal: Communications Biology

    doi: 10.1038/s42003-020-01141-7

    Large-scale DNA data storage in living cells. a The workflow for the manufacture of a mixed culture living cell data storage material. The assembled oligo pool with 10 6 to 10 7 average copies for each oligo was subjected to assembly and then introduced into E. coli cell. A 10 1 to 10 2 average colony number for each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieval and information decoding. b The 0.9% lost oligos in the 1 st passage of the one-fragment assembly (red line) and the 0.56% lost oligos in the 10× deep sequencing reads of the original master pool (blue line) were mapped to the oligo frequency distribution of the original master pool (gray line). c In contrast with previous reported major systems for DNA storage in living cells, including 0.25 kbps by Yachie in 2007, 14.56 bps by Shipman in 2017 and 2.448 kbps by Sun in 2019, the total of 97.728 kbps of DNA for the 509 oligos pool and 2304 Kbps for the 11520 oligos pool stored in a mixed culture of E. coli cells at a cost lower than 0.001$ per base, and the mixed cell storage material could be manufactured within 24 h.
    Figure Legend Snippet: Large-scale DNA data storage in living cells. a The workflow for the manufacture of a mixed culture living cell data storage material. The assembled oligo pool with 10 6 to 10 7 average copies for each oligo was subjected to assembly and then introduced into E. coli cell. A 10 1 to 10 2 average colony number for each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieval and information decoding. b The 0.9% lost oligos in the 1 st passage of the one-fragment assembly (red line) and the 0.56% lost oligos in the 10× deep sequencing reads of the original master pool (blue line) were mapped to the oligo frequency distribution of the original master pool (gray line). c In contrast with previous reported major systems for DNA storage in living cells, including 0.25 kbps by Yachie in 2007, 14.56 bps by Shipman in 2017 and 2.448 kbps by Sun in 2019, the total of 97.728 kbps of DNA for the 509 oligos pool and 2304 Kbps for the 11520 oligos pool stored in a mixed culture of E. coli cells at a cost lower than 0.001$ per base, and the mixed cell storage material could be manufactured within 24 h.

    Techniques Used: Amplification, Plasmid Preparation, Sequencing

    Related Articles

    Polymerase Chain Reaction:

    Article Title: CRISPR-based gene replacement reveals evolutionarily conserved axon guidance functions of Drosophila Robo3 and Tribolium Robo2/3
    Article Snippet: .. Construction of robo3TcRobo2/3 donor plasmid The initial robo3 donor construct was assembled from four PCR fragments via Gibson assembly (New England Biolabs E2611). .. The four fragments were derived from pBluescript (plasmid backbone; primer pair 417–418), the wild-type robo3 genomic locus (5′ and 3′ homology regions; primer pairs 501–502 and 505–506), and the robo3 cDNA (robo3 coding region; primer pair 503–504).

    Clone Assay:

    Article Title: TPXL-1 activates Aurora A to clear contractile ring components from the polar cortex during cytokinesis
    Article Snippet: .. Transgene construction Gibson cloning (E2611; NEB) was used to construct transgenes encoding WT and mNeonGreen tagged TPXL-1 (isoform A) in pCFJ350. ..

    Article Title: Discrete and Structurally Unique Proteins (Tāpirins) Mediate Attachment of Extremely Thermophilic Caldicellulosiruptor Species to Cellulose *
    Article Snippet: .. Tāpirins (Calkro_0844, Calkro_0845) were cloned into the vector pCTCON , using conventional ligation (Calkro_0845) or Gibson Assembly® master mix (Calkro_0844) (New England Biolabs), according to the manufacturer's directions. .. The cloning strategy excluded signal peptides and/or transmembrane domains; oligonucleotide primers used for cloning are listed in .

    Article Title: Temperature-dependent regulation of upstream open reading frame translation in S. cerevisiae
    Article Snippet: .. Generation of C-terminally HA-tagged clones of AGA1 gene Gibson assembly master mix (NEB# E2611) was used to generate C-terminally HA-tagged AGA1 clones [AGA 1-HA (WT), see plasmid #12, Additional file : Table S2]. ..

    Construct:

    Article Title: TPXL-1 activates Aurora A to clear contractile ring components from the polar cortex during cytokinesis
    Article Snippet: .. Transgene construction Gibson cloning (E2611; NEB) was used to construct transgenes encoding WT and mNeonGreen tagged TPXL-1 (isoform A) in pCFJ350. ..

    Article Title: CRISPR-based gene replacement reveals evolutionarily conserved axon guidance functions of Drosophila Robo3 and Tribolium Robo2/3
    Article Snippet: .. Construction of robo3TcRobo2/3 donor plasmid The initial robo3 donor construct was assembled from four PCR fragments via Gibson assembly (New England Biolabs E2611). .. The four fragments were derived from pBluescript (plasmid backbone; primer pair 417–418), the wild-type robo3 genomic locus (5′ and 3′ homology regions; primer pairs 501–502 and 505–506), and the robo3 cDNA (robo3 coding region; primer pair 503–504).

    Plasmid Preparation:

    Article Title: Discrete and Structurally Unique Proteins (Tāpirins) Mediate Attachment of Extremely Thermophilic Caldicellulosiruptor Species to Cellulose *
    Article Snippet: .. Tāpirins (Calkro_0844, Calkro_0845) were cloned into the vector pCTCON , using conventional ligation (Calkro_0845) or Gibson Assembly® master mix (Calkro_0844) (New England Biolabs), according to the manufacturer's directions. .. The cloning strategy excluded signal peptides and/or transmembrane domains; oligonucleotide primers used for cloning are listed in .

    Article Title: Temperature-dependent regulation of upstream open reading frame translation in S. cerevisiae
    Article Snippet: .. Generation of C-terminally HA-tagged clones of AGA1 gene Gibson assembly master mix (NEB# E2611) was used to generate C-terminally HA-tagged AGA1 clones [AGA 1-HA (WT), see plasmid #12, Additional file : Table S2]. ..

    Article Title: CRISPR-based gene replacement reveals evolutionarily conserved axon guidance functions of Drosophila Robo3 and Tribolium Robo2/3
    Article Snippet: .. Construction of robo3TcRobo2/3 donor plasmid The initial robo3 donor construct was assembled from four PCR fragments via Gibson assembly (New England Biolabs E2611). .. The four fragments were derived from pBluescript (plasmid backbone; primer pair 417–418), the wild-type robo3 genomic locus (5′ and 3′ homology regions; primer pairs 501–502 and 505–506), and the robo3 cDNA (robo3 coding region; primer pair 503–504).

    Ligation:

    Article Title: Discrete and Structurally Unique Proteins (Tāpirins) Mediate Attachment of Extremely Thermophilic Caldicellulosiruptor Species to Cellulose *
    Article Snippet: .. Tāpirins (Calkro_0844, Calkro_0845) were cloned into the vector pCTCON , using conventional ligation (Calkro_0845) or Gibson Assembly® master mix (Calkro_0844) (New England Biolabs), according to the manufacturer's directions. .. The cloning strategy excluded signal peptides and/or transmembrane domains; oligonucleotide primers used for cloning are listed in .

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  • 99
    New England Biolabs gibson assembly master mix
    Large-scale DNA data storage in living cells. a The workflow for the manufacture of a <t>mixed</t> culture living cell data storage material. The assembled oligo pool with 10 6 to 10 7 average copies for each oligo was subjected to <t>assembly</t> and then introduced into E. coli cell. A 10 1 to 10 2 average colony number for each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieval and information decoding. b The 0.9% lost oligos in the 1 st passage of the one-fragment assembly (red line) and the 0.56% lost oligos in the 10× deep sequencing reads of the original <t>master</t> pool (blue line) were mapped to the oligo frequency distribution of the original master pool (gray line). c In contrast with previous reported major systems for DNA storage in living cells, including 0.25 kbps by Yachie in 2007, 14.56 bps by Shipman in 2017 and 2.448 kbps by Sun in 2019, the total of 97.728 kbps of DNA for the 509 oligos pool and 2304 Kbps for the 11520 oligos pool stored in a mixed culture of E. coli cells at a cost lower than 0.001$ per base, and the mixed cell storage material could be manufactured within 24 h.
    Gibson Assembly Master Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 411 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gibson assembly master mix/product/New England Biolabs
    Average 99 stars, based on 411 article reviews
    Price from $9.99 to $1999.99
    gibson assembly master mix - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    New England Biolabs hifi dna assembly
    Agarose gel electrophoresis of the three assembled genes (a) The first lane contains the GFP gene assembled from 27 dsDNA fragments showing up at the expected 757 bp length. The second lane contains the kanamycin resistance gene assembled from 28 dsDNA fragments showing up at the expected 953 bp length. The third lane contains the tetracycline resistance gene assembled from 45 dsDNA fragments at the expected 1254 bp length. All assemblies were performed using Gibson Assembly master mix. (b) The same assemblies performed using the NEBuilder <t>HiFi</t> <t>DNA</t> Assembly master mix. The agarose gel is stained with ethidium bromide.
    Hifi Dna Assembly, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 57 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hifi dna assembly/product/New England Biolabs
    Average 99 stars, based on 57 article reviews
    Price from $9.99 to $1999.99
    hifi dna assembly - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Large-scale DNA data storage in living cells. a The workflow for the manufacture of a mixed culture living cell data storage material. The assembled oligo pool with 10 6 to 10 7 average copies for each oligo was subjected to assembly and then introduced into E. coli cell. A 10 1 to 10 2 average colony number for each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieval and information decoding. b The 0.9% lost oligos in the 1 st passage of the one-fragment assembly (red line) and the 0.56% lost oligos in the 10× deep sequencing reads of the original master pool (blue line) were mapped to the oligo frequency distribution of the original master pool (gray line). c In contrast with previous reported major systems for DNA storage in living cells, including 0.25 kbps by Yachie in 2007, 14.56 bps by Shipman in 2017 and 2.448 kbps by Sun in 2019, the total of 97.728 kbps of DNA for the 509 oligos pool and 2304 Kbps for the 11520 oligos pool stored in a mixed culture of E. coli cells at a cost lower than 0.001$ per base, and the mixed cell storage material could be manufactured within 24 h.

    Journal: Communications Biology

    Article Title: A mixed culture of bacterial cells enables an economic DNA storage on a large scale

    doi: 10.1038/s42003-020-01141-7

    Figure Lengend Snippet: Large-scale DNA data storage in living cells. a The workflow for the manufacture of a mixed culture living cell data storage material. The assembled oligo pool with 10 6 to 10 7 average copies for each oligo was subjected to assembly and then introduced into E. coli cell. A 10 1 to 10 2 average colony number for each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieval and information decoding. b The 0.9% lost oligos in the 1 st passage of the one-fragment assembly (red line) and the 0.56% lost oligos in the 10× deep sequencing reads of the original master pool (blue line) were mapped to the oligo frequency distribution of the original master pool (gray line). c In contrast with previous reported major systems for DNA storage in living cells, including 0.25 kbps by Yachie in 2007, 14.56 bps by Shipman in 2017 and 2.448 kbps by Sun in 2019, the total of 97.728 kbps of DNA for the 509 oligos pool and 2304 Kbps for the 11520 oligos pool stored in a mixed culture of E. coli cells at a cost lower than 0.001$ per base, and the mixed cell storage material could be manufactured within 24 h.

    Article Snippet: Then the Gibson Assembly® Master Mix—Assembly (NEB, #E2611) was used according to user’s manual.

    Techniques: Amplification, Plasmid Preparation, Sequencing

    A large-scale DNA data storage in living cell. a) The workflow for the manufacture of mixed culture living cell data storage materials. Oligo pool was assembled with 1E+6⁓7 of average copy of each oligo was subjected to assembly and then transformed into E. coli cell with about 1E+1⁓2 average colony number of each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieve and information decoding. b) the 0.9% dropout oligos in 1 st passaging of one fragment assembly (red line) and the 0.56% dropout oligos in 10x sequencing reads of original master pool (blue line) were mapped to the oligo frequency distribution of original master pool (gray line). c) In comparison with previous reported major systems for DNA storage in living cell including 0.25 kbps by Yachie in 2007, 18.2 bps by Shipman in 2017 and 2.8 kbps by Sun in 2019, totally 97.7 kbps DNA for 509 oligos pool and 2304 kbps for 11520 oligos pool were stored in mixed culture of E. coli cells at cost lower than 1E-4$ per base and mixed cell storage materials could be manufactured within 24 hrs.

    Journal: bioRxiv

    Article Title: Mixed Culture of Bacterial Cell for Large Scale DNA Storage

    doi: 10.1101/2020.02.21.960476

    Figure Lengend Snippet: A large-scale DNA data storage in living cell. a) The workflow for the manufacture of mixed culture living cell data storage materials. Oligo pool was assembled with 1E+6⁓7 of average copy of each oligo was subjected to assembly and then transformed into E. coli cell with about 1E+1⁓2 average colony number of each oligo was obtained and then the cell population could be amplified to large scale in mixed culture for further plasmid retrieve and information decoding. b) the 0.9% dropout oligos in 1 st passaging of one fragment assembly (red line) and the 0.56% dropout oligos in 10x sequencing reads of original master pool (blue line) were mapped to the oligo frequency distribution of original master pool (gray line). c) In comparison with previous reported major systems for DNA storage in living cell including 0.25 kbps by Yachie in 2007, 18.2 bps by Shipman in 2017 and 2.8 kbps by Sun in 2019, totally 97.7 kbps DNA for 509 oligos pool and 2304 kbps for 11520 oligos pool were stored in mixed culture of E. coli cells at cost lower than 1E-4$ per base and mixed cell storage materials could be manufactured within 24 hrs.

    Article Snippet: Then the Gibson Assembly® Master Mix – Assembly (NEB, #E2611) was used according to user’s manual.

    Techniques: Transformation Assay, Amplification, Plasmid Preparation, Passaging, Sequencing

    Polar clearing requires the ability of TPXL-1 to activate Aurora A. (A) Schematics of the protein products of the WT and Aurora A–binding defective (FD) tpxl-1 transgenes. (B) Immunoblots of control (N2) worms and worms expressing TPXL-1 WT or TPXL-1 FD after depletion of endogenous TPXL-1 by RNAi were probed for TPXL-1 and α-tubulin as a loading control. (C) Spindle length calculated by measuring the distance between the centrosomes (Fig. S1 F) is plotted for control (black) and TPXL-1 depleted ( tpxl-1(RNAi) ; gray) embryos and for embryos expressing TPXL-1 WT (green) or TPXL-1 FD (purple) after endogenous TPXL-1 depletion. n = number of embryos. (D) Confocal images of anaphase embryos expressing TPXL-1 WT ::NG ( n = 10) or TPXL-1 FD ::NG ( n = 11) after endogenous TPXL-1 depletion. To visualize TPXL-1::NG on astral microtubules without saturating the aster centers, a gamma of 2.5 was introduced in Photoshop. (E) Time-lapse series of myosin-depleted rga-3/4Δ embryos expressing mKate2::anillin and TPXL-1 WT ( n = 12) or TPXL-1 FD ( n = 8). Embryos were depleted of HCP-4 along with endogenous TPXL-1 to ensure comparable pole separation. (F) Kymographs of the anterior cortex of the embryos in E beginning 180 s after NEBD. (G) Normalized cortical mKate2::anillin fluorescence at the anterior pole; n = number of linescans. (H) Graph plotting the distance between the anterior aster and anterior pole. n = number of embryos. (I) Model illustrating how the activation of Aurora A by TPXL-1 on astral microtubules could generate a diffusible signal that inhibits the accumulation of contractile ring proteins on the polar cortex. All error bars are SEM. Bars, 5 µm.

    Journal: The Journal of Cell Biology

    Article Title: TPXL-1 activates Aurora A to clear contractile ring components from the polar cortex during cytokinesis

    doi: 10.1083/jcb.201706021

    Figure Lengend Snippet: Polar clearing requires the ability of TPXL-1 to activate Aurora A. (A) Schematics of the protein products of the WT and Aurora A–binding defective (FD) tpxl-1 transgenes. (B) Immunoblots of control (N2) worms and worms expressing TPXL-1 WT or TPXL-1 FD after depletion of endogenous TPXL-1 by RNAi were probed for TPXL-1 and α-tubulin as a loading control. (C) Spindle length calculated by measuring the distance between the centrosomes (Fig. S1 F) is plotted for control (black) and TPXL-1 depleted ( tpxl-1(RNAi) ; gray) embryos and for embryos expressing TPXL-1 WT (green) or TPXL-1 FD (purple) after endogenous TPXL-1 depletion. n = number of embryos. (D) Confocal images of anaphase embryos expressing TPXL-1 WT ::NG ( n = 10) or TPXL-1 FD ::NG ( n = 11) after endogenous TPXL-1 depletion. To visualize TPXL-1::NG on astral microtubules without saturating the aster centers, a gamma of 2.5 was introduced in Photoshop. (E) Time-lapse series of myosin-depleted rga-3/4Δ embryos expressing mKate2::anillin and TPXL-1 WT ( n = 12) or TPXL-1 FD ( n = 8). Embryos were depleted of HCP-4 along with endogenous TPXL-1 to ensure comparable pole separation. (F) Kymographs of the anterior cortex of the embryos in E beginning 180 s after NEBD. (G) Normalized cortical mKate2::anillin fluorescence at the anterior pole; n = number of linescans. (H) Graph plotting the distance between the anterior aster and anterior pole. n = number of embryos. (I) Model illustrating how the activation of Aurora A by TPXL-1 on astral microtubules could generate a diffusible signal that inhibits the accumulation of contractile ring proteins on the polar cortex. All error bars are SEM. Bars, 5 µm.

    Article Snippet: Transgene construction Gibson cloning (E2611; NEB) was used to construct transgenes encoding WT and mNeonGreen tagged TPXL-1 (isoform A) in pCFJ350.

    Techniques: Binding Assay, Western Blot, Expressing, Fluorescence, Activation Assay

    Agarose gel electrophoresis of the three assembled genes (a) The first lane contains the GFP gene assembled from 27 dsDNA fragments showing up at the expected 757 bp length. The second lane contains the kanamycin resistance gene assembled from 28 dsDNA fragments showing up at the expected 953 bp length. The third lane contains the tetracycline resistance gene assembled from 45 dsDNA fragments at the expected 1254 bp length. All assemblies were performed using Gibson Assembly master mix. (b) The same assemblies performed using the NEBuilder HiFi DNA Assembly master mix. The agarose gel is stained with ethidium bromide.

    Journal: PLoS ONE

    Article Title: Rational Design of High-Number dsDNA Fragments Based on Thermodynamics for the Construction of Full-Length Genes in a Single Reaction

    doi: 10.1371/journal.pone.0145682

    Figure Lengend Snippet: Agarose gel electrophoresis of the three assembled genes (a) The first lane contains the GFP gene assembled from 27 dsDNA fragments showing up at the expected 757 bp length. The second lane contains the kanamycin resistance gene assembled from 28 dsDNA fragments showing up at the expected 953 bp length. The third lane contains the tetracycline resistance gene assembled from 45 dsDNA fragments at the expected 1254 bp length. All assemblies were performed using Gibson Assembly master mix. (b) The same assemblies performed using the NEBuilder HiFi DNA Assembly master mix. The agarose gel is stained with ethidium bromide.

    Article Snippet: We decided to investigate if Gibson Assembly kit and a recently derived kit called the NEBuilder HiFi DNA Assembly (NEB #E2621) have any inherent limitation on the number of dsDNA fragments that can be assembled at once.

    Techniques: Agarose Gel Electrophoresis, Staining