Diagenode

MeDIP-seq service (Methylated DNA IP Sequencing)

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カタログ番号
フォーマット
G02050000

5mCゲノム全体の研究に、メチル化DNA免疫沈降シーケンシング(MeDIP-Seq)を行います。このアッセイでは、免疫沈降の際の5-mCに対する特異的抗体を利用して、メチル化DNA領域を捕捉します。免疫沈降画分中のメチル化DNAの濃縮は、個々の領域のメチル化状態を評価する配列決定によって確定します。同じ手法を用いて、5-hmChMeDIP-seq)に特異的な抗体を用いることにより、DNAヒドロキシメチル化を研究することも可能です。

DNAメチル化プロファイルの定性分析に優れた方法

    • 100〜500 bpの解像度
    • 低密度および繰り返し頻度の高い領域での5mCカバレッジ
    • 特異性が高く、偏りの少ない配列などに影響を受けない抗体ベースのアプローチ
    • 専門の科学者がハイタッチコミュニケーションで顧客プロジェクトを管理
    • 包括的なサービス - 免疫沈降、ライブラリーの調製、配列決定、分析

 

  • 詳細

    MeDIP and hMeDIP sequencing services include:

    QC of the genomic DNA

    • Measurement of DNA concentration
    • Assessment of  DNA quality

    Preparation of MeDIP-seq/hMeDIP-seq libraries*

    • gDNA shearing on Bioruptor® Pico  
    • Library preparation
    • Immunoprecipitation of methylated or hydroxymethylation DNA using Abs specific to either 5-mC, 5-hmC
    • Library amplification and clean-up  
    • QC of the MeDIP-seq/hMeDIP-seq libraries

    Deep sequencing

    • Samples are sent for sequencing on an Illumina platform  
    • Single-read or paired-end  
    • 50bp to 150bp read length  
    • Greater than or equal to 50M reads per sample
    • Coverage to be defined with customer

    *Full protocol details can be viewed in the MagMeDIP Kit manual

  • バイオインフォマティック解析

    Standard analysis

    Methylation calling:

    Quality check, alignment of sequencing data to reference genome, and identification of methylated or hydroxymethylated regions. Provided files include FastQC reports, alignment files in BAM format, and peak files in BED format.

    Additional analysis on request:  

    Differential methylation analysis:

    Identification of differential methylation or hydroxymethylation between samples based on previously identified MeDIPseq/hMeDIP-seq peaks.  

    Annotation with genomic regions:

    Annotation of MeDIP-seq/hMeDIP-Seq peaks with genomic regions such as introns, exons, enhancers (when available), promoters, intergenic regions.  

    Gene ontology terms analysis:

    Enrichment analysis on gene sets. Gene Ontology terms that are overrepresented in differentially bound regions may indicate the underlying biological processes involved.  

    Pathway analysis:

    Identify biochemical pathways in which genes associated with differentially methylated regions (or individual differentially methylated CpGs) may be overrepresented.  


    Visualization of specific genomic regions:

    Visualization of results (i.e. sequencing data, peaks) at specific genomic regions (e.g. genes, promoters) in publication-ready images (human, mouse, rat).

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  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: MeDIP-seq service (Methylated DNA IP Sequencing) (Diagenode Cat# G02050000). Click here to copy to clipboard.

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イベント

  • Japanese Biochemical Society
    Yokohama
    Sep 18-Sep 20, 2019
  • The 2019 PacBio Long-Read Revolution: Highly Accurate and Affordable SMRT Sequencing
    Atlanta, Georgia
    Sep 19, 2019
  • The 2019 PacBio Long-Read Revolution: Highly Accurate and Affordable SMRT Sequencing
    RTP, North Carolina
    Sep 20, 2019
  • EpiGeneSys
    London
    Sep 22-Sep 24, 2019
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