Post-Bisulfite DNA Library Preparation Kit (For Illumina®) (ab185906)

Overview

  • Product name
    Post-Bisulfite DNA Library Preparation Kit (For Illumina®)
  • Sensitivity
    1 ng
  • Range
    0.5 ng - 1000 ng
  • Assay time
    5h 00m
  • Product overview

    ab185906 is a complete set of optimized reagents to prepare a DNA library -- after successful bisulfite conversion -- for various Illumina® platform-based bisulfite sequencing (bisulfite-seq) assays, such as whole genome bisulfite sequencing (WGBS), oxidative bisulfite sequencing (oxBs-seq), reduced representative bisulfite sequencing (RRBS), and other bisulfite-based next generation sequencing applications. The optimized protocol and components of the kit allow both non-barcoded (singleplexed) and barcoded (multiplexed) DNA libraries to be quickly constructed using sub-nanogram input concentrations of DNA since the DNA is first bisulfite-converted and then used for library preparation.

     

    Illumina® is a registered trademark of Illumina, Inc.

  • Notes

    Several methods, including whole genome bisulfite sequencing (WGBS) and reduced representation bisulfite sequencing (RRBS), are currently used for genome-wide DNA methlyation analysis. These methods convert unmethylated cytosines to uracil while 5-methylcytosines remain unchanged by the bisulfite treatment. This allows epigenetic differences to become genetic differences, which can be subsequently detected via sequencing at the single-based resolution level and on a genome-wide scale. However, practical use with such current methods are not ideal as they (1) need large amounts of DNA (>1 µg) as input material, which is difficult to prepare from limited biological samples such as tumor biopsy, early embyros, embyronic tissues, and circulating DNA; (2) require DNA to first be sheared and then ligated to adapters, followed by bisulfite conversion (post-ligation bisulfite conversion), which causes substantial amounts of DNA fragments contained in the adapter-DNA fragment constructs to be broken and thereby forms mono-tagged templates that become removed during library enrichment; and (3) are time-consuming (2 days). The Post-Bisulfite DNA Library Preparation Kit (For Illumina®) is designed to overcome these weaknesses.

Properties

  • Storage instructions
    Please refer to protocols.
  • Components 12 tests 24 tests
    10X dA-Tailing Buffer 1 x 40µl 1 x 80µl
    10X End Repair Buffer 1 x 40µl 1 x 80µl
    2X HiFi PCR Master Mix 1 x 160µl 1 x 320µl
    2X Ligation Buffer 1 x 250µl 1 x 500µl
    5X Conversion Buffer 1 x 50µl 1 x 100µl
    Adaptors (50 μM) 1 x 15µl 1 x 30µl
    Conversion Enzyme Mix 1 x 15µl 1 x 30µl
    Conversion Primer 1 x 26µl 1 x 52µl
    Elution Buffer 1 x 1ml 1 x 2ml
    End Repair Enzyme Mix 1 x 25µl 1 x 50µl
    Klenow Fragment (3’-5’ exo-) 1 x 15µl 1 x 30µl
    MQ Binding Beads 1 x 1.8ml 1 x 3.6ml
    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

  • Size distribution of library fragments as demonstrated by a post-bisulfite DNA library constructed using ab185906 Post-Bisulfite DNA Library Preparation Kit (For Illumina®) from 10 ng of input DNA.

Protocols

References

ab185906 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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