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EpiNext High-Sensitivity Bisulfite-Seq Kit (Illumina)
For streamlined bisulfite sequencing NGS library preparation from low input DNA

EpiNext 高靈敏度 DNA 甲基化定序建庫套組


DNA methylation occurs by the covalent addition of a methyl group (CH3) at the 5-carbon of the cytosine ring, resulting in 5-methylcytosine (5-mC). DNA methylation is essential in regulating gene expression in nearly all biological processes including development, growth, and differentiation. Alterations in DNA methylation have been demonstrated to cause a change in gene expression. For example, hypermethylation leads to gene silencing or decreased gene expression while hypomethylation activates genes or increases gene expression. Aberrant DNA methylation is also associated with pathogenesis of diseases such as cancer, autoimmune disorders, and schizophrenia. Thus, genome-wide analysis of DNA methylation could provide valuable information for discovering epigenetic markers used for disease diagnosis, and potential targets used for therapeutics.

The EpiNext™ High-Sensitivity Bisulfite-Seq Kit (Illumina) is designed to bisulfite-convert DNA and prepare an Illumina-based library for bisulfite sequencing, all in one kit. Intended applications include whole genome (WGBS), oxidative (oxBS-seq), reduced representation (RRBS), and other bisulfite-next generation sequencing. The optimized protocol and components of the kit allow subnanogram amounts of DNA to be bisulfite converted and fragmented simultaneously followed by quick non-barcoded (singleplexed) and barcoded (multiplexed) library construction in less than 7 hours.

實驗原理

This kit includes all reagents required for successfully preparing a library directly using bisulfite DNA generated from a tiny amount of input DNA. In this preparation, DNA is simultaneously bisulfite converted and fragmented to the appropriate length during the bisulfite process. The bisulfite-treated DNA, which is in single stranded form, is then simultaneously converted to dsDNA and adaptor ligated. The ligated fragments are size selected and purified using MQ binding beads, which allows for quick and precise size selection of DNA. Size-selected DNA fragments are amplified using a high-fidelity PCR Mix which ensures maximum yields from minimum amounts of starting material and provides highly accurate amplification of library DNA with low error rates and minimal bias.

Workflow of the EpiNext™ High-Sensitivity Bisulfite-Seq Kit (Illumina).

The EpiNext™ High-Sensitivity Bisulfite-Seq Kit (Illumina) was used to bisulfite convert DNA and prepare a library to be sequencing on an Illumina HiSeq 2500. Data tracks were aligned to a 30 kb region of chromosome 1 and showed unmethylated (blue) and methylated (red) areas and the corresponding gene regions in treated and control samples.



Size distribution of library fragments. Post-bisulfite DNA libraries were prepared from human placenta DNA using the EpiNext™ High-Sensitivity Bisulfite-Seq Kit (Illumina): A: 10 ng; B: 50 ng.

產品列表

貨號 產品 數量
P-1056A-12 EpiNext High-Sensitivity Bisulfite-Seq Kit (Illumina) 高靈敏亞硫酸鹽定序試劑套組 (組) 廠牌:Epigentek
P-1056A-24 EpiNext High-Sensitivity Bisulfite-Seq Kit (Illumina) 高靈敏亞硫酸鹽定序試劑套組 (組) 廠牌:Epigentek

EpiNext High-Sensitivity Bisulfite-Seq Kit (Illumina)

  • Innovative method : simultaneous bisulfite conversion and size-appropriate DNA fragmentation.
  • Fast and streamlined procedure : completed within 6 hours and 30 minutes
  • Complete conversion (>99%)
  • High sensitivity and efficiency : input DNA to be as low as 0.2 ng
  • Extremely convenient : The kit contains all required components for each step of DNA library preparation.
  • Minimal bias : Ultra HiFi amplification
  • Broad sample suitability
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