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nebuilder hifi dna assembly  (New England Biolabs)


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    Structured Review

    New England Biolabs nebuilder hifi dna assembly
    Nebuilder Hifi Dna Assembly, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 9454 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nebuilder+hifi+dna+assembly+master+mix/NEBuilder+HiFi+DNA+Assembly+Master+Mix/bio_rxiv__64898__2026__10__15__759383-62-4-4
    Average 99 stars, based on 9454 article reviews
    nebuilder hifi dna assembly - by Bioz Stars, 2026-11
    99/100 stars

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    Related Articles

    other:

    Article Title: Decoding how RNase E membrane targeting controls RNA degradosome activity in Pseudomonas aeruginosa
    Article Snippet: NEBuilder HiFi DNA assembly Master Mix , New England Biolabs , Cat# E2621L.

    Amplification:

    Article Title: Haematopoietic Cells Deficient for Calreticulin or Expressing Its Oncogenic Variants Maintain Functional Antigen Presentation
    Article Snippet: .. Primers were ordered from Sigma–Aldrich; fragments were amplified by PrimeSTAR Max DNA Polymerase (Takara), assembled by NEBuilder HiFi DNA Assembly Master Mix (NEB), and constructs were validated by Sanger sequencing. ..

    Construct:

    Article Title: Haematopoietic Cells Deficient for Calreticulin or Expressing Its Oncogenic Variants Maintain Functional Antigen Presentation
    Article Snippet: .. Primers were ordered from Sigma–Aldrich; fragments were amplified by PrimeSTAR Max DNA Polymerase (Takara), assembled by NEBuilder HiFi DNA Assembly Master Mix (NEB), and constructs were validated by Sanger sequencing. ..

    Article Title: Myopathy-causing variants of PYROXD1 converge on impaired RNA ligation
    Article Snippet: .. Constructs were assembled from gel-purified PCR products or from synthesized DNA fragments using NEBuilder HiFi DNA Assembly Master Mix (NEB, E262), according to the manufacturer’s instructions. .. PCR fragments were amplified using Q5 Hot Start High-Fidelity DNA Polymerase (NEB, M0493), either using cDNA or existing lab-internal plasmids as template.

    DNA Sequencing:

    Article Title: Haematopoietic Cells Deficient for Calreticulin or Expressing Its Oncogenic Variants Maintain Functional Antigen Presentation
    Article Snippet: .. Primers were ordered from Sigma–Aldrich; fragments were amplified by PrimeSTAR Max DNA Polymerase (Takara), assembled by NEBuilder HiFi DNA Assembly Master Mix (NEB), and constructs were validated by Sanger sequencing. ..

    Synthesized:

    Article Title: Solute selectivity and functional divergence of aquaporins in reef-building corals
    Article Snippet: Full-length cDNAs encoding Acropora millepora Aqps (AmiAqps) were chemically synthesized ( Table S1 ; Eurofins Genomics, Tokyo, Japan) with 15 bp terminal sequences homologous to the ends of the linearized pGEMHE vector ( ). .. The synthesized cDNAs were subcloned into pGEMHE using the NEBuilder HiFi DNA Assembly Master Mix (New England Biolabs, Ipswich, MA, USA). .. All constructs were verified by Sanger sequencing using an Applied Biosystems 3730xl DNA Analyzer (Thermo Fisher Scientific, Waltham, MA, USA).

    Article Title: Myopathy-causing variants of PYROXD1 converge on impaired RNA ligation
    Article Snippet: .. Constructs were assembled from gel-purified PCR products or from synthesized DNA fragments using NEBuilder HiFi DNA Assembly Master Mix (NEB, E262), according to the manufacturer’s instructions. .. PCR fragments were amplified using Q5 Hot Start High-Fidelity DNA Polymerase (NEB, M0493), either using cDNA or existing lab-internal plasmids as template.

    Gel Purification:

    Article Title: A Third-generation Lentiviral Vector System for Flow Virometry-based Analysis of HIV-1 Env and Other Viral Envelope Proteins
    Article Snippet: .. Following DpnI digestion and gel purification of all PCR products, the fluorophore coding sequences were inserted between the duplicated HIV-1 protease cleavage sites using the NEBuilder HiFi DNA Assembly Master Mix according to the manufacturer’s instructions. ..

    Polymerase Chain Reaction:

    Article Title: A Third-generation Lentiviral Vector System for Flow Virometry-based Analysis of HIV-1 Env and Other Viral Envelope Proteins
    Article Snippet: .. Following DpnI digestion and gel purification of all PCR products, the fluorophore coding sequences were inserted between the duplicated HIV-1 protease cleavage sites using the NEBuilder HiFi DNA Assembly Master Mix according to the manufacturer’s instructions. ..

    Article Title: Myopathy-causing variants of PYROXD1 converge on impaired RNA ligation
    Article Snippet: .. Constructs were assembled from gel-purified PCR products or from synthesized DNA fragments using NEBuilder HiFi DNA Assembly Master Mix (NEB, E262), according to the manufacturer’s instructions. .. PCR fragments were amplified using Q5 Hot Start High-Fidelity DNA Polymerase (NEB, M0493), either using cDNA or existing lab-internal plasmids as template.

    Clone Assay:

    Article Title: Engineering a Modular Bacterial Galactose Oxidase through CBM Recombination and Second-Shell Substitutions
    Article Snippet: For all PCR products, the DNA template was digested with 10 U of DpnI (Thermo Scientific) at 37 °C for 6 h, followed by purification using the Illustra GFX PCR DNA kit (GE Healthcare). .. The truncated PsGalOx variants and CBM genes were cloned into pET-15b using NEBuilder HiFi DNA Assembly Master Mix, with a 1:10 vector:insert ratio for 1 h at 50 °C. .. The ligated mixture was dialyzed through a 0.025 μm nitrocellulose membrane (Merck Millipore) and then used to transform the electrocompetent E. coli DH5α strain.

    Molecular Cloning:

    Article Title: Engineering a Modular Bacterial Galactose Oxidase through CBM Recombination and Second-Shell Substitutions
    Article Snippet: For all PCR products, the DNA template was digested with 10 U of DpnI (Thermo Scientific) at 37 °C for 6 h, followed by purification using the Illustra GFX PCR DNA kit (GE Healthcare). .. The truncated PsGalOx variants and CBM genes were cloned into pET-15b using NEBuilder HiFi DNA Assembly Master Mix, with a 1:10 vector:insert ratio for 1 h at 50 °C. .. The ligated mixture was dialyzed through a 0.025 μm nitrocellulose membrane (Merck Millipore) and then used to transform the electrocompetent E. coli DH5α strain.

    Plasmid Preparation:

    Article Title: Engineering a Modular Bacterial Galactose Oxidase through CBM Recombination and Second-Shell Substitutions
    Article Snippet: For all PCR products, the DNA template was digested with 10 U of DpnI (Thermo Scientific) at 37 °C for 6 h, followed by purification using the Illustra GFX PCR DNA kit (GE Healthcare). .. The truncated PsGalOx variants and CBM genes were cloned into pET-15b using NEBuilder HiFi DNA Assembly Master Mix, with a 1:10 vector:insert ratio for 1 h at 50 °C. .. The ligated mixture was dialyzed through a 0.025 μm nitrocellulose membrane (Merck Millipore) and then used to transform the electrocompetent E. coli DH5α strain.

    Article Title: One‐Step Induction of Oogonia‐Like and Pro‐Spermatogonia‐Like Cells From Porcine Pluripotent Stem Cells
    Article Snippet: .. The purified target gene fragments were assembled with the linearized vector backbone using NEBuilder HiFi DNA Assembly Master Mix (E2621L) via homologous recombination to complete the construction of TRE3G‐pSOX17, TRE3G‐pTFAP2C, and TRE3G‐pBLIMP1 vectors. ..

    Purification:

    Article Title: One‐Step Induction of Oogonia‐Like and Pro‐Spermatogonia‐Like Cells From Porcine Pluripotent Stem Cells
    Article Snippet: .. The purified target gene fragments were assembled with the linearized vector backbone using NEBuilder HiFi DNA Assembly Master Mix (E2621L) via homologous recombination to complete the construction of TRE3G‐pSOX17, TRE3G‐pTFAP2C, and TRE3G‐pBLIMP1 vectors. ..

    Homologous Recombination:

    Article Title: One‐Step Induction of Oogonia‐Like and Pro‐Spermatogonia‐Like Cells From Porcine Pluripotent Stem Cells
    Article Snippet: .. The purified target gene fragments were assembled with the linearized vector backbone using NEBuilder HiFi DNA Assembly Master Mix (E2621L) via homologous recombination to complete the construction of TRE3G‐pSOX17, TRE3G‐pTFAP2C, and TRE3G‐pBLIMP1 vectors. ..



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    New England Biolabs hifi assembly
    A. Schematic of paired Perturb-seq experimental design with summaries of each experiment’s sgRNA expression strategy and recovered cell populations. B. Expression percentile of target TFs in cells containing each targeting sgRNA and all control sgRNAs. Marginal histogram shows distribution of expression percentiles for all target genes in all perturbed cells (blue) or all control cells (gray). C. ChromVAR motif enrichment of perturbed subpopulations (grouped by target TF) with at least one differential motif ( q < 0.1, Kolmogorov-Smirnov test) across all motifs of target TFs. D. UMAP visualization of joint embedding of gene expression and chromatin phenotypes for all perturbed subpopulations, grouped by sgRNA. Points are colored by target TF in alphabetical order with a continuous color scale, such that homologs are assigned similar colors. Inset UMAPs show average activity of motifs in JASPAR FOX and AP-1 motif clusters. E. Energy distance calculated on normalized components of gene expression (GEX) PCA embedding and chromatin (ATAC) spectral embedding for all targets with at least 20 cells. Colored points represent targets with significant energy distance difference from control cells ( q < 0.01, permutation test, Benjamini-Hochberg correction) in at least one modality. TFs are colored as in  . F. On-target ChromVAR motif enrichment vs. number of high-confidence newly expressed genes for all targets with at least 20 cells. High-confidence newly expressed genes are unexpressed in control cells, expressed in at least 10% of target cells, and associated with at least one concordant (differentially accessible) ATAC peak. Colored points indicate target TFs selected for dual-guide Perturb-seq screen; coloring as in  . G. Schematic of priming between illustrative TFs A and B, in which A “primes” B by opening B motifs after activation. H. Priming relationships among 50 TFs selected for dual-guide Perturb-seq screen. A single chord is drawn between primed TF pairs and colored by the TF whose motif is opened, with colors as in  . Chords between bidirectional pairs are colored by the TF whose motif has the larger z-score.
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    Image Search Results


    Journal: iScience

    Article Title: Decoding how RNase E membrane targeting controls RNA degradosome activity in Pseudomonas aeruginosa

    doi: 10.1016/j.isci.2026.117717

    Figure Lengend Snippet:

    Article Snippet: NEBuilder HiFi DNA assembly Master Mix , New England Biolabs , Cat# E2621L.

    Techniques: Produced, Virus, Sequencing, Recombinant, Yeast, Saline, Reverse Transcription, Software, Sterility

    A. Schematic of paired Perturb-seq experimental design with summaries of each experiment’s sgRNA expression strategy and recovered cell populations. B. Expression percentile of target TFs in cells containing each targeting sgRNA and all control sgRNAs. Marginal histogram shows distribution of expression percentiles for all target genes in all perturbed cells (blue) or all control cells (gray). C. ChromVAR motif enrichment of perturbed subpopulations (grouped by target TF) with at least one differential motif ( q < 0.1, Kolmogorov-Smirnov test) across all motifs of target TFs. D. UMAP visualization of joint embedding of gene expression and chromatin phenotypes for all perturbed subpopulations, grouped by sgRNA. Points are colored by target TF in alphabetical order with a continuous color scale, such that homologs are assigned similar colors. Inset UMAPs show average activity of motifs in JASPAR FOX and AP-1 motif clusters. E. Energy distance calculated on normalized components of gene expression (GEX) PCA embedding and chromatin (ATAC) spectral embedding for all targets with at least 20 cells. Colored points represent targets with significant energy distance difference from control cells ( q < 0.01, permutation test, Benjamini-Hochberg correction) in at least one modality. TFs are colored as in  . F. On-target ChromVAR motif enrichment vs. number of high-confidence newly expressed genes for all targets with at least 20 cells. High-confidence newly expressed genes are unexpressed in control cells, expressed in at least 10% of target cells, and associated with at least one concordant (differentially accessible) ATAC peak. Colored points indicate target TFs selected for dual-guide Perturb-seq screen; coloring as in  . G. Schematic of priming between illustrative TFs A and B, in which A “primes” B by opening B motifs after activation. H. Priming relationships among 50 TFs selected for dual-guide Perturb-seq screen. A single chord is drawn between primed TF pairs and colored by the TF whose motif is opened, with colors as in  . Chords between bidirectional pairs are colored by the TF whose motif has the larger z-score.

    Journal: bioRxiv

    Article Title: Engineered regulatory states resolve context-dependent genetics in macular degeneration

    doi: 10.64898/2026.10.09.758047

    Figure Lengend Snippet: A. Schematic of paired Perturb-seq experimental design with summaries of each experiment’s sgRNA expression strategy and recovered cell populations. B. Expression percentile of target TFs in cells containing each targeting sgRNA and all control sgRNAs. Marginal histogram shows distribution of expression percentiles for all target genes in all perturbed cells (blue) or all control cells (gray). C. ChromVAR motif enrichment of perturbed subpopulations (grouped by target TF) with at least one differential motif ( q < 0.1, Kolmogorov-Smirnov test) across all motifs of target TFs. D. UMAP visualization of joint embedding of gene expression and chromatin phenotypes for all perturbed subpopulations, grouped by sgRNA. Points are colored by target TF in alphabetical order with a continuous color scale, such that homologs are assigned similar colors. Inset UMAPs show average activity of motifs in JASPAR FOX and AP-1 motif clusters. E. Energy distance calculated on normalized components of gene expression (GEX) PCA embedding and chromatin (ATAC) spectral embedding for all targets with at least 20 cells. Colored points represent targets with significant energy distance difference from control cells ( q < 0.01, permutation test, Benjamini-Hochberg correction) in at least one modality. TFs are colored as in . F. On-target ChromVAR motif enrichment vs. number of high-confidence newly expressed genes for all targets with at least 20 cells. High-confidence newly expressed genes are unexpressed in control cells, expressed in at least 10% of target cells, and associated with at least one concordant (differentially accessible) ATAC peak. Colored points indicate target TFs selected for dual-guide Perturb-seq screen; coloring as in . G. Schematic of priming between illustrative TFs A and B, in which A “primes” B by opening B motifs after activation. H. Priming relationships among 50 TFs selected for dual-guide Perturb-seq screen. A single chord is drawn between primed TF pairs and colored by the TF whose motif is opened, with colors as in . Chords between bidirectional pairs are colored by the TF whose motif has the larger z-score.

    Article Snippet: Vector pEM070 ( , top) was created from pEM040 (Addgene #224898) by digesting with BsiWI and BbsI, removing the eGFP coding sequence, and replacing with a gBlock (IDT) encoding eGFP with a C-terminal fusion of the KASH domain from human SYNE2 (isoform 1, amino acids 6804-6885) via HiFi assembly (NEB #E2621).

    Techniques: Expressing, Control, Gene Expression, Activity Assay, Activation Assay, Expression Vector