The BisulFlash™ DNA Modification Kit is a complete set

of optimized buffers and reagents to perform DNA modification using a next

 generation DNA bisulfite conversion technology developed by Epigentek.

 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 bisulfite DNA 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 > 99.99%.
• 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, real time MS-PCR, methylation microarray, and pyrosequencing. Additionally, based on its ability for a complete cytosine conversion, it is specifically suitable for next generation methylation sequencing/pyrosequencing.

Comparative Overview of Commercial Kits
Data was obtained through actual use of the kit, customer feedback, or information provided by the suppliers datasheet or website.

Fig. 2. 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 DNA-specific and converted DNA-specific primers (β-actin, 110 bps),

 respectively. A: real time PCR; B: end-point PCR. The BisulFlash™ kit

 treated DNA was completely converted, and no unconverted DNA in the

treated samples was determined after 45 cycles.

 DNA at various amounts (0.2 ng-200 ng) were

converted using the BisulFlash™ DNA Modification Kit. 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 BisulFlash™ DNA Modification Kit.

A: real time PCR; B: starting DNA amount-CT value curve.