Overview

  • Product name

    High-Sensitivity DNA Library Preparation Kit (For Illumina®)
  • Assay time

    1h 30m
  • Product overview

    High Sensitivity DNA Library Preparation KIt (ab185905) is a complete set of optimized reagents to prepare a DNA library from very small amount of samples for use in next-generation sequencing applications. This kit is suitable for preparing a DNA library using sub-nanogram amounts of DNA input for next generation sequencing applications using an Illumina sequencer. These applications include genomic DNA-seq, ChIP-seq, MeDIP/hMeDIP-seq, traditional bisulfite-seq, and targeted re-sequencing. The optimized protocol and components of the kit allow both non-barcoded (singleplexed) and barcoded (multiplexed) DNA libraries to be constructed quickly with reduced bias.

     

    Starting Materials
    Starting materials can include fragmented dsDNA isolated from various tissue or cell samples, dsDNA enriched from a ChIP reaction, MeDIP/hMeDIP reaction, or exon capture. DNA should be relatively free of RNA because large fractions of RNA will impair end repair and dA-tailing, resulting in reduced ligation capabilities. The input amount of DNA can be from 0.2 ng to 100 ng. For optimal preparation, the input amount should be 10 ng to 50 ng.

     

     

    Illumina® is a registered trademark of Illumina, Inc.

  • Notes

    DNA library preparation is a critical step for next generation sequencing (NGS). To generate accurate sequencing data in NGS, the prepared library DNA should be sufficient in yield and of high quality. As NGS technology is continuously improving, DNA library preparation is required to be optimized accordingly. Most of the currently used methods are time-consuming, expensive, inconvenient, and specifically need large amounts of DNA. These factors result in a DNA library preparation which cannot be used for biological samples with limited amounts of starting material such as tumor biopsy, early embryos, and embryonic tissues and circulating DNA. In addition, the amount of DNA enriched by ChIP or MeDIP/hmeDIP is often at low or sub-nanogram levels which causes insufficient DNA library yields. To address this issue, Abcam offers ab185905, the High-Sensitive DNA Library Preparation Kit (For Illumina®).

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 12 tests 24 tests
    10X End Polishing Buffer 1 x 30µl 1 x 60µl
    2X HiFi PCR Master Mix 1 x 160µl 1 x 320µl
    2X Ligation Buffer 1 x 250µl 1 x 500µl
    Adaptors (50 μM) 1 x 15µl 1 x 30µl
    Elution Buffer 1 x 1ml 1 x 2ml
    End Polishing Enhancer 1 x 13µl 1 x 26µl
    End Polishing Enzyme Mix 1 x 13µl 1 x 26µl
    MQ Binding Beads 1 x 1.6ml 1 x 3.2ml
    Primer I (10 μM) 1 x 15µl 1 x 30µl
    Primer U (10 μM) 1 x 15µl 1 x 30µl
    T4 DNA Ligase 1 x 15µl 1 x 30µl
  • Research areas

Images

  • Human placenta DNA was sheared to around 300 bps in peak size and 0.2 ng of DNA was used for DNA library preparation using High Sensitivity DNA Library Preparation Kit (ab185905).

Protocols

References

This product has been referenced in:

  • Pasquier C  et al. The Mapping of Predicted Triplex DNA:RNA in the Drosophila Genome Reveals a Prominent Location in Development- and Morphogenesis-Related Genes. G3 (Bethesda) 7:2295-2304 (2017). Read more (PubMed: 28515050) »
See 1 Publication for this product

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