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  • NEBNext® Fast DNA Fragmentation & Library Prep Set for Ion Torrent™

    Description






    The NEBNext® Fast DNA Fragmentation & Library Prep Set for Ion Torrent™ contains enzymes and buffers in convenient master mix formulations that are ideally suited for sample preparation for next-generation sequencing on the Ion Torrent Sequencer (Life Technologies, Inc.). Each of these components must pass rigorous quality control standards and are lot controlled, both individually and as a set of reagents.

    Functional Validation
    Each set of reagents is functionally validated together through construction and sequencing of a DNA library on the Ion Torrent platform.
    DNA Library Preparation Workflow for Ion Torrent

    Representative Results from Libraries Constructed Using NEBNext for Ion Torrent
    0.5 µg of DNA from 3 different genomes with varying GC content were used to construct 200 bp and 400 bp libraries using the NEBNext Fast DNA Fragmentation and Library Prep Set for Ion Torrent, analyzed by the Agilent® Bioanalyzer®.

    Representative Ion Torrent Run Report
    A typical Ion Torrent run report for libraries made from E. coli (K12 MG1655 strain) genomic DNA using NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent. A. Alignment Summary. 72 Mb of data was generated, with an average genome coverage of 15.5 X, from approximately 0.5 Million 200 bp reads. B. Read length histogram from 200 bp run.

    Figure 1: Relative size distribution of Fragmented End Repaired DNA as seen using the Bioanalyzer® 2100 (Agilent Technologies, Inc.).
    1 µg of E. coli DNA was fragmented and end repaired for 20 minutes at 25°C, followed by 10 minutes at 70°C.
    Figure 2: Final Library size distribution using AMPure XP Bead Size Selection.
    Figure 3: Final Library Size distribution using E-Gel Size Selection.

    Lot Control

    The lots provided are managed separately and qualified by additional functional validation. Individual reagents undergo standard enzyme activity and quality control assays, and also meet stringent criteria in the additional quality controls listed on each individual component page

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    NEBNext DNA Fragmentation Master Mix
    T4 DNA Ligase
    NEBNext DNA Library Primers for Ion Torrent™1X
    TE Buffer0.1X
    NEBNext Stop Buffer
    Magnesium Chloride (MgCl2) (200 mM)
    NEBNext DNA Library Adaptors for Ion Torrent
    NEBNext DNA Fragmentation Reaction Buffer10X
    T4 DNA Ligase Buffer for Ion Torrent™10X
    NEBNext High-Fidelity 2X PCR Master Mix-202X
    NEBNext® Q5® Hot Start HiFi PCR Master Mix-20
    Bst 2.0 WarmStart® DNA Polymerase-20

    Properties and Usage

    Materials Required but not Supplied

    • Agencourt® AMPure® XP Beads (Beckman Coulter, Inc., #A63881) or PCR Column Purification Kit (Qiagen or other)
    • 80% ethanol (freshly prepared)
    • Size selection materials [E-Gel® (Life Technologies, Inc.) or AMPURE XP Beads etc.]

    Storage Temperature

    -20°C

    FAQs

    1. What sequencing platform can I use the NEBNext remove DNA Fragmentation & Library Prep Sets for Ion Torrent reagents for?
    2. What type of starting material can be used with NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent?
    3. How much starting material do I need to use when preparing libraries using the NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent?
    4. Does the NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent work well for GC-rich DNA and AT-rich DNA?
    5. Can I use the NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent for preparing RNA?
    6. What methods for size selection can be used with Ion Torrent Library Preparation Kits?
    7. Are these reagents available in a customized format or bulk format?
    8. What do I do if my fragmented DNA is too short?
    9. The peak on my electropherogram trace is broader than I would like. How can I make the size range more narrow?
    10. Do I really need to vortex the fragmentation reactions, as described in the protocol? Will pipetting up and down work just as well?
    11. What methods for size selection can be used with Ion Torrent Library Preparation Kits?
    12. What do I do if my DNA is not being fragmented?
    13. Are DNA adaptors or oligonucleotides included in the NEBNext products?
    14. Are NEBNext® products validated in Next Generation Sequencing workflows?

    Protocols

    1. Fragmentation and End Repair of DNA Protocol (E6285)
    2. Preparation of Adaptor Ligated DNA (E6285)
    3. Cleanup of Adaptor Ligated DNA (E6285)
    4. Size Selection (E6285)
    5. PCR Amplification of Adaptor Ligated DNA (E6285)
    6. Clean Up of Amplified Library (E6285)

    Manuals

    The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name these document files: manual[Catalog Number].

    Safety Data Sheet

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.