EpiSeeker Bisulfite Modification Kit - Whole Cell (ab117126)

Overview

  • Product nameEpiSeeker Bisulfite Modification Kit - Whole CellSee all DNA modification kits ...
  • Sample type
    Saliva, Urine, Tissue, Adherent cells, Suspension cells
  • Assay time
    3h 00m
  • Species reactivity
    Reacts with: Human
    Predicted to work with: A wide range of mammals
  • Product overview

    DNA methylation is a covalent modification of the cytosine ring at the 5' position of a CpG dinucleotide which leads to epigenetic inactivation of genes when found in 5'-CpG-3'dinucleotides within promoters or in the first exon of genes. It is well demonstrated that DNA methylation plays an important role in the regulation of gene expression, tumorigenesis, and other genetic and epigenetic diseases.

    Most methods for DNA methylation detection require a prior bisulfite-based DNA modification. Bisulfite-based DNA modification is used to discrimate between cytosine and methylated cytosine, in which DNA is treated with bisulfite salt to convert cytosine residues to uracil in a single-stranded DNA, while methylated cytosine remains the same.

    EpiSeeker Bisulfite Modification Kit - Whole Cell (ab117126) allows the user to perform DNA methylation analysis and modify DNA directly from cells or tissues within only 3 hours. ab117126 is specifically designed for DNA methylation research using minute amounts of starting materials including cells cultured in 96-well/384-well plates, tissue section samples, microdissection samples, tissue biopsy and early embryonic cells/oocytes.

    Modified DNA is eluted and suitable for real time MS-PCR as well as for all techniques currently used for the analysis of DNA methylation; including conventional MS-PCR, bisulfite sequencing, pyrosequencing, and methylation microarray.

    Not sure if this is the right kit for you? Check out our bisulfite modification guide for more information.

  • Notes

    NOTE: If you use ab117126 for MSP, the numbers of PCR cycles should be greater than 45.

    The kit can be stored at room temperature (20-22°C) away from light (with the exception of component EBM1 (Digestion Powder) for 6 months from the shipping date.

    EBM1 should be stored at –20°C, or stored at 4°C as soon as it is dis-solved in EBM2 (up to 6 months).

    Each vial of EBM4 can be used for 10 sample treatments.

    The prepared EBM4/EBM5/EBM6 solution should be used immediately, unless it is stored at –20°C away from light (up to one week).

    Frozen EBM4/EBM5/EBM6 solution must be thawed at room temperature and vortexed for 2 minutes prior to use.

  • Tested applicationsFunctional Studiesmore details

Properties

  • Storage instructionsPlease refer to protocols.
  • Components 40 tests 80 tests
    EBM1 (Digestion Powder) 1 vial 1 vial
    EBM2 (Digestion Solution) 1 x 0.1ml 1 x 0.2ml
    EBM3 (Cell Collection Buffer) 1 x 1ml 1 x 2ml
    EBM4 (DNA Modification Powder) 4 vials 8 vials
    EBM5 (DNA Modification Buffer) 1 x 5ml 1 x 10ml
    EBM6 (Balance Buffer) 1 x 0.4ml 1 x 0.8ml
    EBM7 (DNA Binding Buffer) 1 x 14ml 1 x 28ml
    EBM8 (Modified DNA Elution) 1 x 1ml 1 x 2ml
    F-Collection Tube 1 x 40 units 1 x 80 units
    F-Spin Column 1 x 40 units 1 x 80 units
  • Research Areas

    Applications

    Our Abpromise guarantee covers the use of ab117126 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    Functional Studies Use at an assay dependent concentration. Minimum starting material: 100-20000 cells / 1-100µg tissue / 0.2-2mm2 tissue section sample. Optimal starting material: 500-5000 cells / 5-20µg tissue / 0.5-1mm2 tissue section.

    EpiSeeker Bisulfite Modification Kit - Whole Cell images

    • MCF-7 cells were modified using ab117126. 10 µl of modified DNA were eluted and 2 µl of elution were used in real time PCR. A pair of primers and a probe designed to amplify both methylated and unmethylated alleles of ß-actin was used.

    Protocols

    References for EpiSeeker Bisulfite Modification Kit - Whole Cell (ab117126)

    ab117126 has not yet been referenced specifically in any publications.

    Product Wall

    After bisulfite the DNA will be in ssDNA form and can not be directly used for ligation with illumina's adaptor. We recommend to use ab117127 (enhancer program) to bisulfite modifying DNA after ligation with Illumina's adaptor as indicated by Illumina ...

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    Thank you very for having patience!
    If use extracted DNA, the procedure can be processed starting from step 3 with slight modification:

    3. Add 1 ml of EBM5 to 1 vial of EBM4, followed by adding 60 µl of EBM6 to create the EBM4/EBM5/EBM6 so...

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    Thank you for contacting us.
    The kit can be used with tissue as low as 1 microgram and with paraffin embedded tissue sections. The procedure is given in attached booklet;
    http://www.abcam.com/ps/products/117/ab117126/documents/ab117126%20EpiS...

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    Thank you for your patience regarding this shipment of the digestion buffer for ab117126. I am very sorry for the delay. I have issued a free of charge replacement.You should receive this tomorrow.

    Please note that this free of charge replace...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"