• My NEB
  • Print
  • PDF
  • CpG Methylated NIH 3T3 Mouse Genomic DNA

    Description

    NIH 3T3 (mouse embryonic fibroblast cell line) genomic DNA that was enzymatically methylated with CpG Methylase (M.SssI), suitable as a positive control in the study of CpG dinucleotide methylation.

    Advantages and Features

    Applications

    • A positive 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 Methylation, Bisulfite Treatment):
      CpG methylated genomic DNA is bisulfite converted and tested in a PCR reaction with primers specific to CpG methylated PTEN or Rb promoter DNA and compared to a PCR reaction with primers that anneal to unmethylated PTEN or Rb DNA. Only the methylated-specific primer stes generate the appropriate sized PCR product.
    • Functional Test (Genomic DNA Methylation, Radioactivity Incorporation):
      CpG methylated genomic DNA is tested in a reaction containing radiolabelled SAM and CpG methylase. After incubation the percentage of labeled genomic DNA is determined by radioactive incorporation

    Notes

    1. A 260/280 Ratio: 1.90

    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.