nebnext ultra ii dna library prep kit for illumina  (New England Biolabs)


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  • 99
    Name:
    NEBNext Ultra II DNA Library Prep Kit for Illumina
    Description:
    NEBNext Ultra II DNA Library Prep Kit for Illumina 96 rxns
    Catalog Number:
    e7645l
    Price:
    2245
    Size:
    96 rxns
    Category:
    DNA Template Preparation for PCR
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    New England Biolabs nebnext ultra ii dna library prep kit for illumina
    NEBNext Ultra II DNA Library Prep Kit for Illumina
    NEBNext Ultra II DNA Library Prep Kit for Illumina 96 rxns
    https://www.bioz.com/result/nebnext ultra ii dna library prep kit for illumina/product/New England Biolabs
    Average 99 stars, based on 15048 article reviews
    Price from $9.99 to $1999.99
    nebnext ultra ii dna library prep kit for illumina - by Bioz Stars, 2020-05
    99/100 stars

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    1) Product Images from "RELACS nuclei barcoding enables high-throughput ChIP-seq"

    Article Title: RELACS nuclei barcoding enables high-throughput ChIP-seq

    Journal: Communications Biology

    doi: 10.1038/s42003-018-0219-z

    RELACS workflow. Overview of the RELACS method. The protocol facilitates barcoding multiple cell populations, which can be pooled and investigated for multiple epitopes within the same run. The method starts by isolating nuclei from a pool of formaldehyde-fixed cells, using sonication to reduce cell type dependency 5 ( a ). The nuclear membrane is permeabilized to allow entrance of enzymes, followed by DNA barcodes. Restriction endonucleases with a high frequency of recognition sites are used to fragment chromatin—in this work CviKI-1 was used ( b ). Nuclei are washed to remove active restriction enzymes ( c ). Hairpin adapters harboring barcodes are ligated to both the ends of the fragmented chromatin inside the nuclei. The barcoding has been tested using 100–500,000 nuclei without the need to change protocol conditions ( d ). Cell populations marked with specific barcodes are pooled ( e ), concentrated and lysed using SDS at low concentration and short sonication to release chromatin into solution ( f ). Chromatin is split and incubated with the antibodies of interest ( g ). After ChIP washes and DNA purification (not illustrated), only DNA that harbors nuclei barcodes at both ends is PCR amplified to complete library construction. PCR amplification appends an Illumina barcode onto mark each fragment ( h ). Sequenced libraries are demultiplexed by Illumina barcode, to retrieve ChIP information, and then nuclear barcode, to identify the initial cell population ( i ). The RELACS protocol is very fast and ChIP-seq libraries can be generated for hundreds of samples within three days
    Figure Legend Snippet: RELACS workflow. Overview of the RELACS method. The protocol facilitates barcoding multiple cell populations, which can be pooled and investigated for multiple epitopes within the same run. The method starts by isolating nuclei from a pool of formaldehyde-fixed cells, using sonication to reduce cell type dependency 5 ( a ). The nuclear membrane is permeabilized to allow entrance of enzymes, followed by DNA barcodes. Restriction endonucleases with a high frequency of recognition sites are used to fragment chromatin—in this work CviKI-1 was used ( b ). Nuclei are washed to remove active restriction enzymes ( c ). Hairpin adapters harboring barcodes are ligated to both the ends of the fragmented chromatin inside the nuclei. The barcoding has been tested using 100–500,000 nuclei without the need to change protocol conditions ( d ). Cell populations marked with specific barcodes are pooled ( e ), concentrated and lysed using SDS at low concentration and short sonication to release chromatin into solution ( f ). Chromatin is split and incubated with the antibodies of interest ( g ). After ChIP washes and DNA purification (not illustrated), only DNA that harbors nuclei barcodes at both ends is PCR amplified to complete library construction. PCR amplification appends an Illumina barcode onto mark each fragment ( h ). Sequenced libraries are demultiplexed by Illumina barcode, to retrieve ChIP information, and then nuclear barcode, to identify the initial cell population ( i ). The RELACS protocol is very fast and ChIP-seq libraries can be generated for hundreds of samples within three days

    Techniques Used: Sonication, Concentration Assay, Incubation, Chromatin Immunoprecipitation, DNA Purification, Polymerase Chain Reaction, Amplification, Generated

    Related Articles

    Methylation Sequencing:

    Article Title: Designer epigenome modifiers enable robust and sustained gene silencing in clinically relevant human cells
    Article Snippet: .. For next generation bisulfite sequencing, libraries were constructed from PCR Amplicons using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, E7645) and quantified using the ddPCR Library Quantification Kit for Illumina TruSeq (Biorad, #186–3040). .. All samples were sequenced on an Illumina MiSeq platform with a MiSeq Reagent Micro Kit v2, 300 cycles (Illumina, MS-103-1002).

    Multiplex Assay:

    Article Title: Isolation and genome-wide characterization of cellular DNA:RNA triplex structures
    Article Snippet: .. DNA libraries were prepared from at least three independent experiments from HeLa S3 cells using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB) and NEBNext Multiplex Oligos for Illumina (NEB). .. Single-end sequencing was performed on Illumina NextSeq 500 platform.

    Modification:

    Article Title: Fluorescent amplification for next generation sequencing (FA-NGS) library preparation
    Article Snippet: .. Conclusions Here we present a useful modification to conventional NGS library preparation workflows, FA-NGS, which was successfully incorporated into Illumina Nextera XT and NEBNext Ultra II DNA library preparation. .. We demonstrate the FA-NGS workflow ease of use with fewer overall steps than conventional library workflows, as well as an MCA QC test to confirm successful library construction before sequencing.

    Ligation:

    Article Title: Single-stranded DNA library preparation from highly degraded DNA using T4 DNA ligase
    Article Snippet: .. The second double-stranded method was originally proposed by Illumina ( ) and relies on T/A-overhang ligation of a fork-like adapter, or, in the implementation of New England Biolabs’ NEBNext Ultra II DNA Library Prep Kit for Illumina used here, a bell-shaped adapter (Figure ). .. The Illumina method enables directional ligation and a greater degree of simplification, e.g. by combining some or all of the reactions without intermittent purification steps.

    Article Title: RELACS nuclei barcoding enables high-throughput ChIP-seq
    Article Snippet: .. Ligation of barcodes to A-tailed chromatin was performed using components from NEBNext Ultra II DNA library preparation kit, with the following modifications: per sample 15 µL of ligation master mix and 0.5 µL of ligation enhancer were added, followed by 15 min incubation at 30 °C and 15 min incubation at 20 °C. ..

    Next-Generation Sequencing:

    Article Title: Fluorescent amplification for next generation sequencing (FA-NGS) library preparation
    Article Snippet: .. Conclusions Here we present a useful modification to conventional NGS library preparation workflows, FA-NGS, which was successfully incorporated into Illumina Nextera XT and NEBNext Ultra II DNA library preparation. .. We demonstrate the FA-NGS workflow ease of use with fewer overall steps than conventional library workflows, as well as an MCA QC test to confirm successful library construction before sequencing.

    Construct:

    Article Title: Designer epigenome modifiers enable robust and sustained gene silencing in clinically relevant human cells
    Article Snippet: .. For next generation bisulfite sequencing, libraries were constructed from PCR Amplicons using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, E7645) and quantified using the ddPCR Library Quantification Kit for Illumina TruSeq (Biorad, #186–3040). .. All samples were sequenced on an Illumina MiSeq platform with a MiSeq Reagent Micro Kit v2, 300 cycles (Illumina, MS-103-1002).

    Purification:

    Article Title: Avoidance of Trinucleotide Corresponding to Consensus Protospacer Adjacent Motif Controls the Efficiency of Prespacer Selection during Primed Adaptation
    Article Snippet: .. Amplicons containing plasmid prespacers and extended CRISPR arrays were gel purified and used to create Illumina sequencing libraries with an NEBNext Ultra II DNA library preparation kit with U5 barcoding. .. High-throughput sequencing of amplicons was conducted on MiniSeq or HiSeq Illumina machines using the 2 × 150 paired-end mode.

    Polymerase Chain Reaction:

    Article Title: Designer epigenome modifiers enable robust and sustained gene silencing in clinically relevant human cells
    Article Snippet: .. For next generation bisulfite sequencing, libraries were constructed from PCR Amplicons using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, E7645) and quantified using the ddPCR Library Quantification Kit for Illumina TruSeq (Biorad, #186–3040). .. All samples were sequenced on an Illumina MiSeq platform with a MiSeq Reagent Micro Kit v2, 300 cycles (Illumina, MS-103-1002).

    Incubation:

    Article Title: Solid-phase enzyme catalysis of DNA end repair and 3′ A-tailing reduces GC-bias in next-generation sequencing of human genomic DNA
    Article Snippet: .. Similar to the method using PKT, the protocols of NEBNext Ultra II DNA Library Prep Kit and Kapa Hyper Prep Kit adopt high temperature incubation at 65 °C (Fig. ). .. The protocol of Illumina TruSeq DNA PCR-free LT Library Preparation Kit includes a 5 min incubation at 70 °C following 3′ A-tailing for 30 min at 37 °C (Fig. ).

    Article Title: RELACS nuclei barcoding enables high-throughput ChIP-seq
    Article Snippet: .. Ligation of barcodes to A-tailed chromatin was performed using components from NEBNext Ultra II DNA library preparation kit, with the following modifications: per sample 15 µL of ligation master mix and 0.5 µL of ligation enhancer were added, followed by 15 min incubation at 30 °C and 15 min incubation at 20 °C. ..

    CRISPR:

    Article Title: Avoidance of Trinucleotide Corresponding to Consensus Protospacer Adjacent Motif Controls the Efficiency of Prespacer Selection during Primed Adaptation
    Article Snippet: .. Amplicons containing plasmid prespacers and extended CRISPR arrays were gel purified and used to create Illumina sequencing libraries with an NEBNext Ultra II DNA library preparation kit with U5 barcoding. .. High-throughput sequencing of amplicons was conducted on MiniSeq or HiSeq Illumina machines using the 2 × 150 paired-end mode.

    Sequencing:

    Article Title: Avoidance of Trinucleotide Corresponding to Consensus Protospacer Adjacent Motif Controls the Efficiency of Prespacer Selection during Primed Adaptation
    Article Snippet: .. Amplicons containing plasmid prespacers and extended CRISPR arrays were gel purified and used to create Illumina sequencing libraries with an NEBNext Ultra II DNA library preparation kit with U5 barcoding. .. High-throughput sequencing of amplicons was conducted on MiniSeq or HiSeq Illumina machines using the 2 × 150 paired-end mode.

    Plasmid Preparation:

    Article Title: Avoidance of Trinucleotide Corresponding to Consensus Protospacer Adjacent Motif Controls the Efficiency of Prespacer Selection during Primed Adaptation
    Article Snippet: .. Amplicons containing plasmid prespacers and extended CRISPR arrays were gel purified and used to create Illumina sequencing libraries with an NEBNext Ultra II DNA library preparation kit with U5 barcoding. .. High-throughput sequencing of amplicons was conducted on MiniSeq or HiSeq Illumina machines using the 2 × 150 paired-end mode.

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