Next Generation Sequencing
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  • DNA Library Preparation

    Library preparation for the major next generation sequencing platforms requires the ligation of specific adaptor oligos to fragments of the DNA to be sequenced. First, DNA is fragmented to the optimal length determined by the downstream platform. Because DNA fragmentation does not result in homogeneous, blunt-ended fragments, end repair is needed to ensure that each molecule is free of overhangs, and contains 5’ phosphate and 3’ hydroxyl groups. Libraries to be used in blunt-ended adaptor ligation, including Ion Torrent™ or SOLiD™ 4 library construction, can be used directly in the ligation step. For Illumina® libraries and some libraries intended for the 454™ platform, incorporation of a non-templated deoxyadenosine 5’-monophosphate (dAMP) onto the 3’ end of blunted DNA fragments, a process known as dA-tailing, is necessary. dA-tails prevent concatamer formation during downstream ligation steps, and enable DNA fragments to be ligated to adaptors with complementary dT-overhangs. The desired adaptor ligated DNA size for Illumina, SOLiD and Ion Torrent platforms can be selected via gel electrophoresis before amplification by the polymerase chain reaction (PCR). New England Biolabs supplies reagents for DNA library preparation for the leading sequencing platforms. For more information, choose the DNA Library Construction Workflow tab below.

    Ion Torrent™ and SOLiD™ 4 are trademarks owned by Life Technologies, Inc.
    Illumina® is a registered trademark of Illumina, Inc.
    454™ is a trademark of Roche.
    1. Tips for Optimizing DNA Inputs in NGS Library Construction

      Optimizing DNA inputs for NGS library prep is an important step, if you want to ensure a high-quality library. Start by following these tips.

    2. NEBNext for Core Lab Facilities

      Are you running a core sequencing facility? Watch as Eileen shares a few reasons that NEBNext products are particularly well suited to sequencing core labs.

    3. Why Choose NEBNext Reagents for NGS Library Prep?

      NEBNext products are a great choice for your NGS library preparation needs. In this video, Fiona shares a few reasons why.

    4. 12 Quick Tips for NGS Library Preparation

      Using beads to clean up your DNA prior to NGS library prep can be quick and easy, if you follow these few, simple tips.

    FAQs for DNA Library Preparation

    Protocols for DNA Library Preparation

    DNA Library Construction Workflow