Fast Bisulfite Conversion Kit (ab117127)
Key features and details
- Assay time: 30 min
Overview
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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
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Storage instructions
Store at room temperature. Please refer to protocols. -
Components 50 tests Capture Solution 1 x 15ml Conversion Enhancer 5 vials Conversion Mix Solution 1 x 6ml Denaturation Enhancer 1 x 600µl Desulphonation Solution 1 x 60µl Elution Solution 1 x 1ml F-Collection Tube 1 pack F-Spin Column 1 pack -
Research areas
Images
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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.
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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.