Overview

  • Product name

    Fast Bisulfite Conversion Kit
    See all DNA modification kits
  • Assay time

    0h 30m
  • Product overview

    Fast DNA modification Kit (ab117127) allows DNA denaturation and bisulfite conversion to be processed at the same time so the complete procedure can be performed in only 30 minutes. Furthermore, it prevents more than 90% of DNA loss, completely converting unmethylated cytosine into uracil.


    Traditional methods involve a separate denaturation step followed by a subsequent sodium bisulfite DNA conversion step - but with ab117127, DNA denaturation status is concurrently sustained throughout the entire bisulfite DNA conversion process. ab117127 is suitable for MS-PCR, real time MS-PCR, bisulfite sequencing, pyrosequencing and methylation microarray.


    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. 


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

Properties

Images

  • Effective DNA Protection: Fully methylated human genomic DNA at various amounts (0.2 ng-200 ng) were converted using the ab117127. 1 µl of 20 µl eluate was used for real time qPCR and a pair of primers was used to amplify converted DNA. As little as 0.2 ng DNA is sufficient for bisulfite conversion using the ab117127. A: real time PCR; B: starting DNA amount-CT value curve.
  • Complete Cytosine Conversion: 200 ng of genomic DNA isolated from 3 cancer cell lines was treated with ab117127. Next, the unconverted and converted DNA in each treated sample were determined using unconverted DNA-specific and converted DNA-specific primers (ß-actin, 110 bps), respectively. A: real time PCR; B: end-point PCR. The ab117127 treated DNA was completely converted, and no unconverted DNA in the treated samples was determined after 45 cycles.

Protocols

References

This product has been referenced in:

  • Kint S  et al. Evaluation of bisulfite kits for DNA methylation profiling in terms of DNA fragmentation and DNA recovery using digital PCR. PLoS One 13:e0199091 (2018). Read more (PubMed: 29902267) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

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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Answer

Thank you for contacting us.

I am happy to let you know that the EpiSeeker Fast DNA modification Kitab117127is also suitable for bisulfite sequencing and pyrosequencing. Here are several papers related to use of this kit for bisulfite sequencing:

Martínez-Macías, MI. et. al. (February 2012). A DNA 3' phosphatase functions in active DNA demethylation in Arabidopsis. Mol Cell. 45 (3):357-70.

Yang, J. et. al. (November 2011). Stem Cell Gene SALL4 Suppresses Transcription through Recruitment of DNA Methyltransferases.J Biol Chem.Epub ahead of print.

Geyer, K. et. al. (August 2011). Cytosine methylation regulates oviposition in the pathogenic blood fluke Schistosoma mansoni. Nat Commun. 2:424.

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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