Diagenode

MagMeDIP qPCR Kit

目录号
格式
价格
C02010021
(mc-magme-048)
48 rxns (IP)
$710.00
其他格式

Sensitive tumour detection and classification using plasma cell-free DNA methylomes
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Preparation of cfMeDIP-seq libraries for methylome profiling of plasma cell-free DNA
Read the method

Perform MeDIP (Methylated DNA Immunoprecipitation) followed by qPCR to estimate DNA methylation status of your sample using 5-methylcytosine antibody. Our kit contains high quality reagents to get the highest enrichment of methylated DNA with an optimized user-friendly protocol.

To perform an NGS compatible MeDIP-seq assay, you will need the MagMeDIP-seq Package.

Additionally, the development of Diagenode’s MagMeDIP qPCR Kit has been largely inspired by the reproducibility standards reached using the SX-8G IP-Star® Automated System. If you are interested in automating this protocol, please check our Auto MagMeDIP qPCR Kit.

Characteristics

  • Generate highly consistent results with internal controls in 24h
  • Minimize error with many reagents in 1 tube
  • Optimized purification (DIB - DNA isolation buffer)
  • Allows direct correlation between IP’d material & methylation status

MagMeDIP kit validated using Bioruptor

Figure 1. Immunoprecipitation results obtained with Diagenode MagMeDIP qPCR Kit. MeDIP assays were performed manually using DNA from HeLa, colon and blood cells with the MagMeDIP qPCR kit (Diagenode). The IP was performed with Methylated and Unmethylated spike-in controls included in the kit, together with the human DNA sample controls. The DNA was isolated/purified using DIB. Afterwards, qPCR was performed using the primer pairs included in this kit.

  • MagMeDIP workflow

    DNA methylation occurs primarily as 5-methylcytosine (5-mC), and the Diagenode MagMeDIP Kit takes advantage of a specific antibody targeting this 5-mC to immunoprecipitate methylated DNA, which can be thereafter directly analyzed by qPCR or Next-Generation Sequencing (NGS).

    How it works

    In brief, after the cell collection and lysis, the genomic DNA is extracted, sheared, and then denatured. In the next step the antibody directed against 5 methylcytosine and antibody binding beads are used for immunoselection and immunoprecipitation of methylated DNA fragments. Then, the IP’d methylated DNA is isolated and can be used for any subsequent analysis as qPCR, amplification, hybridization on microarrays or next generation sequencing.

    5-methylcytosine
  • MeDIP-qPCR

    The kit MagMeDIP contains all reagents necessary for a complete MeDIP-qPCR workflow. Two MagMeDIP protocols have been validated: for manual processing as well as for automated processing, using the Diagenode’s IP-Star Compact Automated System (please refer to the kit manual).

    • Complete kit including DNA extraction module, IP antibody and reagents, DNA isolation buffer
    • Quality control of the IP: due to methylated and unmethylated DNA spike-in controls and their associated qPCR primers
    • Easy to use with user-friendly magnetic beads and rack
    • Highly validated protocol
    • Automated protocol supplied
    Methylated DNA Immunoprecipitation

    Figure 1. Immunoprecipitation results obtained with Diagenode MagMeDIP Kit

    MeDIP assays were performed manually using 1 µg or 50 ng gDNA from blood cells with the MagMeDIP kit (Diagenode). The IP was performed with the Methylated and Unmethylated spike-in controls included in the kit, together with the human DNA samples. The DNA was isolated/purified using DIB. Afterwards, qPCR was performed using the primer pairs included in this kit.

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  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: MagMeDIP qPCR Kit (Diagenode Cat# C02010021). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Pre-diagnosis plasma cell-free DNA methylome profiling up to sevenyears prior to clinical detection reveals early signatures of breast cancer
    Cheng N. et al.
    Profiling of cell-free DNA (cfDNA) has been well demonstrated to be a potential non-invasive screening tool for early cancer detection. However, limited studies have investigated the detectability of cfDNA methylation markers that are predictive of cancers in asymptomatic individuals. We performed cfDNA methylation ...

    Cell-free multi-omics analysis reveals tumor status-informativesignatures in gastrointestinal cancer patients’ plasma
    Tao Y. et al.
    During cancer development, host’s tumorigenesis and immune signals are released to and informed by circulating molecules, like cell-free DNA (cfDNA) and RNA (cfRNA) in blood. However, these two kinds of molecules are still not systematically compared in gastrointestinal cancer. Here, we profiled 4 types of cel...

    Methylation and expression of glucocorticoid receptor exon-1 variants andFKBP5 in teenage suicide-completers.
    Rizavi H. et al.
    A dysregulated hypothalamic-pituitary-adrenal (HPA) axis has repeatedly been demonstrated to play a fundamental role in psychiatric disorders and suicide, yet the mechanisms underlying this dysregulation are not clear. Decreased expression of the glucocorticoid receptor (GR) gene, which is also susceptible to epigen...

    Bridging biological cfDNA features and machine learning approaches.
    Moser T. et al.
    Liquid biopsies (LBs), particularly using circulating tumor DNA (ctDNA), are expected to revolutionize precision oncology and blood-based cancer screening. Recent technological improvements, in combination with the ever-growing understanding of cell-free DNA (cfDNA) biology, are enabling the detection of tumor-speci...

    Neonatal inflammation increases hippocampal KCC2 expression throughmethylation-mediated TGF-β1 downregulation leading to impairedhippocampal cognitive function and synaptic plasticity in adult mice.
    Rong J. et al.
    The mechanisms by which neonatal inflammation leads to cognitive deficits in adulthood remain poorly understood. Inhibitory GABAergic synaptic transmission plays a vital role in controlling learning, memory and synaptic plasticity. Since early-life inflammation has been reported to adversely affect the GABAergic syn...

    Impact of FecB Mutation on Ovarian DNA Methylome inSmall-Tail Han Sheep.
    Xie L. et al.
    UNLABELLED: Booroola fecundity (FecB) gene, a mutant of bone morphogenetic protein 1B (BMPR-1B) that was discovered in Booroola Merino, was the first prolificacy gene identified in sheep related to increased ovulation rate and litter size. The mechanism of FecB impact on reproduction is unclear. METHODS: In this stu...

    Longitudinal monitoring of cell-free DNA methylation in ALK-positivenon-small cell lung cancer patients.
    Janke Florian et al.
    BACKGROUND: DNA methylation (5-mC) signals in cell-free DNA (cfDNA) of cancer patients represent promising biomarkers for minimally invasive tumor detection. The high abundance of cancer-associated 5-mC alterations permits parallel and highly sensitive assessment of multiple 5-mC biomarkers. Here, we performed ...

    Cerebrospinal fluid methylome-based liquid biopsies for accuratemalignant brain neoplasm classification.
    Zuccato Jeffrey A et al.
    BACKGROUND: Resolving the differential diagnosis between brain metastases (BM), glioblastomas (GBM), and central nervous system lymphomas (CNSL) is an important dilemma for the clinical management of the main three intra-axial brain tumor types. Currently, treatment decisions require invasive diagnostic surgical bio...

    Consistent DNA Hypomethylations in Prostate Cancer.
    Araúzo-Bravo M.J. et al.
    With approximately 1.4 million men annually diagnosed with prostate cancer (PCa) worldwide, PCa remains a dreaded threat to life and source of devastating morbidity. In recent decades, a significant decrease in age-specific PCa mortality has been achieved by increasing prostate-specific antigen (PSA) screening and i...

    Cell-free DNA methylation-defined prognostic subgroups in small celllung cancer identified by leukocyte methylation subtraction
    Ul Haq Sami et al.
    Small cell lung cancer (SCLC) methylome is understudied. Here, we comprehensively profile SCLC using cell-free methylated DNA immunoprecipitation followed by sequencing (cfMeDIP-seq). Cell-free DNA (cfDNA) from plasma of 74 SCLC patients pre-treatment and from 20 non-cancer participants, genomic DNA (gDNA) from peri...

    The cell-free DNA methylome captures distinctions between localized andmetastatic prostate tumors.
    Chen Sujun et al.
    Metastatic prostate cancer remains a major clinical challenge and metastatic lesions are highly heterogeneous and difficult to biopsy. Liquid biopsy provides opportunities to gain insights into the underlying biology. Here, using the highly sensitive enrichment-based sequencing technology, we provide analysis of 60 ...

    A SOX2-engineered epigenetic silencer factor represses the glioblastomagenetic program and restrains tumor development.
    Benedetti V. et al.
    Current therapies remain unsatisfactory in preventing the recurrence of glioblastoma multiforme (GBM), which leads to poor patient survival. By rational engineering of the transcription factor SOX2, a key promoter of GBM malignancy, together with the Kruppel-associated box and DNA methyltransferase3A/L catalytic dom...

    mTORC1 is required for epigenetic silencing during β-cell functionalmaturation.
    Ni Qicheng et al.
    OBJECTIVE: The mechanistic target of rapamycin comple×1 (mTORC1) is a key molecule that links nutrients, hormones, and growth factors to cell growth/function. Our previous studies have shown that mTORC1 is required for β-cell functional maturation and identity maintenance; however, the underlying mechanis...

    Detection of ovarian cancer using plasma cell-free DNA methylomes.
    Lu Huaiwu et al.
    BACKGROUND: Ovarian cancer (OC) is a highly lethal gynecologic cancer, and it is hard to diagnose at an early stage. Clinically, there are no ovarian cance