BisulFlash DNA Bisulfite Modification Kit (30 Minute DNA Bisulfite Conversion Kit for DNA Methylation Analysis)

Product Description

The BisulFlash™ DNA Modification Kit is a complete set of optimized buffers and reagents to perform DNA modification using an expedited DNA bisulfite conversion technology developed by Epigentek, geared specifically for methylation-specific PCR (MSP) as well as post-bisulfite based NGS applications. Through a proprietary composition which allows DNA denaturation and bisulfite conversion to be processed at the same time, the complete procedure is reduced to only 30 minutes. Furthermore, it prevents more than 90% of DNA loss, completely converting unmethylated cytosine into uracil.

Perfecting Bisulfite Conversion

Traditional methods involve a separate denaturation step followed by a subsequent sodium bisulfite DNA conversion step -- but with the BisulFlash™ method, DNA denaturation status is concurrently sustained throughout the entire DNA bisulfite conversion process. This breakthrough approach enables the DNA conversion process to be significantly faster with higher conversion efficiency and accuracy. We continue to innovate with the development of the new BisulFlash™ kit by identifying four critical components of bisulfite conversion:

• Speed: Reduce the entire procedure to as short as 30 minutes without any reagent setup time.
• Efficiency: Completely convert unmethylated cytosine into uracil -- modified DNA > 99.9%.
• DNA Protection: Protect against DNA degradation of which more than 90% of DNA loss can be prevented, allowing for greater recovery.
• Sensitivity: Start with the lowest amount of input DNA for modification -- only 0.2 ng or just 50 cells.

The convenient ready-to-use, DNA conversion mix solution and single temperature incubation along with the features mentioned above allow for true perfection in bisulfite conversion. The BisulFlash™ DNA Modification Kit is suitable for MS-PCR/MSP and real time MSP.

Bisulfite Sequencing Option

The BisulFlash™ kit includes a special step made specifically for better sequencing results and is 100% compatible with all Illumina® platforms. For ideal NGS applications, we recommend combining this technique with our EpiNext Post-Bisulfite Library Preparation Kit.

Fig. 1. High accuracy of DNA conversion is achieved by the BisulFlash™ DNA Modification Kit. 50 ng of genomic DNA methylated in all CpG sites by DNA methylases was treated with the BisulFlash™ DNA Modification Kit. Converted DNA was amplified by real time qPCR using primers for multiple promoters containing numerous CpG sites and then directly sequenced.

Product Comparison

Data was obtained through actual use of the kit, customer feedback, or information provided by the supplier's datasheet or website.

Complete Cytosine Conversion

200 ng of genomic DNA isolated from 3 cancer cell lines was treated with the BisulFlash DNA Modification Kit. Next, the unconverted and converted DNA in each treated sample were determined using unconverted DNAspecific and converted DNA-specific primers (β-actin, 110 bp), respectively.

Fig.2. Graph: Real-time PCR; Gel image: End-point PCR. The treated DNA was completely converted, and no unconverted DNA was determined after 45 cycles.