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  • 5-Aza-dc Treated Jurkat Genomic DNA

    Description

    Genomic DNA purified from human male Jurkat (human acute T-cell leukemia) cells that are treated with 5-aza-2-deoxycytidine (5-Aza-dc), suitable as a negative control in the study of CpG dinucleotide methylation in the genome.

    Advantages and Features

    Applications

    • A negative control for Methylation-Specific PCR (MSP) (2), Bisulfite sequencing, Methylation-sensitive Single-Nucleotide Primer Extension (Ms-SNuPE), Combined Bisulfite Restriciton Analysis (COBRA), Bisulfite treatment and PCR-Single-Strand Conformation Polymorphism Analysis (Bisulfite-PCR-SSCP/BiPS).

    Properties and Usage

    Storage Temperature

    -20°C

    Quality Control

    Quality Assurance Statement

    • Purified free of contaminating proteins and RNA.

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Functional Test (Genomic DNA Demethylation, Bisulfite Treatment):

      5-Aza-dc treated genomic DNA is bisulfite converted and tested in a PCR reaction with primers specific to the CpG methylated IGS ribosomal DNA. The percentage of demethylated CpG dinucleotides is determined by DNA sequencing of the appropriate sized DNA products.

    Notes

    1. A260/280 Ratio: 1.85
    2. The potent methyltransferase inhibitor (MTI) 5-aza-2-deoxycytidine (5-Aza-dc) (Decitabine, Dacogen) causes growth arrest, differentiation, and/or apoptosis of many cell types in vitro and in vivo. The genomic DNA derived from cells treated with this drug exhibit some lower molecular weight smearing when visualized on a 0.8% agarose gel. Significant (up to 70%) genome-wide CpG demethylation was confirmed by bisulfite sequencing of IGS ribosomal DNA (rDNA).

    References

    1. Sambrook, J. and Russell, D. (2001). Molecular Cloning: A Laboratory Manual. (3rd ed.), (pp. 6.4-6.12). Cold Spring Harbor: Cold Spring Ha.
    2. Herman, J. G. and Baylin, S. B. (1996). U.S. Patent No. 5,786,146, Johns Hopkins University School of Medicine.
    3. Frommer, M., et. al. (1992). PNAS USA. 89, 1827-1831.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.