Bisulfite Conversion Kit - Whole Cell (ab117126)

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Overview

  • Product name

    Bisulfite Conversion Kit - Whole Cell
    See 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: 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.

     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

     

     

     

Properties

Images

  • Functional Studies - Bisulfite Modification Kit - Whole Cell (ab117126)
    Functional Studies - Bisulfite Modification Kit - Whole Cell (ab117126)
    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

ab117126 has not yet been referenced specifically in any publications.

Publishing research using ab117126? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Question

Can you please explain how this kit works i. e. is the DNA still double stranded after treatment and can you ligate Illumina adapters to it? Thanks,

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Answer

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 Bisulfiote-Seq instruction. If using ab1171266, EpiSeeker Bisulfite Modification Kit - Whole Cell, start from step 4 (page 13) of the protocol to bisuflfite modifying DNA after ligation with Illumina's adaptor.

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Question

Inquiry: i have paraffin embedded tissue, but in eppendorf tube,so: 1. I can proceed my bisulfate conversion on these sample (after deparaffinization with xylene then ethanol directely on these sample)? 2. if i have DNA already my DNA extracted, how i can proceed my Bisulfate conversion by your product (code: 117126)? 3. Our paraffin embedded tissue are very old so we get very low DNA yield....did your product (117126) modify this low DNA amount without DNA degradation and loss? Thank you....

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Answer

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 solution. Vortex until solution is clear or saturated (about 2 minutes).
4. Add DNA sample into PCR tube and then add 110 µl of the mixed EBM4/EBM5/EBM6 solution to each PCR tube containing the sample. Place the tube in a thermal cycler (with heated lid) and program the thermal cycler as followed:
95°C for 6minutes
65°C for 90 minutes
95°C for 3 minutes
Modified sample can then be held at 25°C in the thermal cycler up to 4 hours without loss of performance.
Then goes to the next steps of the protocol.
I hope this will help.

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Question

Product code: 117126
Inquiry: i have paraffin embedded tissue, but in eppendorf tube,so: 1. I can proceed my bisulfate conversion on these sample (after deparaffinization with xylene then ethanol directely on these sample)? 2. if i have DNA already my DNA extracted, how i can proceed my Bisulfate conversion by your product (code: 117126)? 3. Our paraffin embedded tissue are very old so we get very low DNA yield....did your product (117126) modify this low DNA amount without DNA degradation and loss? Thank you....

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Answer

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;
https://www.abcam.com/ps/products/117/ab117126/documents/ab117126%20EpiSeeker%20Bisulfite%20Modification%20Kit%20Whole%20Cell%20(Website).pdf
This kit does not involve DNA isolation and purification step so the DNA loss will be minimal with generation of high yield of modified DNA.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

Expecting Digestion buffer for kit ab117126.

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Answer

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 replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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