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 OptionThe 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.
Comparative Overview of Commercial KitsData was obtained through actual use of the kit, customer feedback, or information provided by the supplier's datasheet or website.
Principle & ProcedureAs a next generation bisulfite conversion tool, the BisulFlash™ DNA Modification Kit contains all reagents required for an ultra-fast bisulfite conversion on a DNA sample. With the ready-to-use conversion mix solution, DNA denaturation status is sustained throughout the entire bisulfite DNA conversion process. This proprietary solution allows the bisulfite reagents to rapidly convert all cytosine to uracil with negligible methylcytosine conversion. The unique DNA protection reagents contained in the mix can prevent the chemical and thermophilic degradation of DNA in the bisulfite treatment. The non-toxic DNA capture solution enables DNA to tightly bind to the column filter, thus DNA cleaning can be carried out on the column to effectively remove residual bisulfite and salts. Rapid ResultsOnly 30 minutes are required for the entire BisulFlash™ procedure -- from converting your sample DNA to pure bisulfite modified DNA. That is significantly faster than any currently used kits (2-8 h) or homebrew methods (>16 h). The BisulFlash™ kit provides everything required for a successful bisulfite conversion and DNA cleanup in the shortest time with the fewest steps possible. Perfect DNA ConversionEach reaction with the BisulFlash™ kit can use 0.2 ng – 1 µg of DNA. For optimal conversion, the DNA amount is 50-200 ng. The novel procedure and proprietary ready-to-use DNA conversion mix solution allow DNA denaturation and bisulfite conversion to occur at the same time and enable all cytosines to be converted to uracil (>99.9%), while 5-methylcytosine remains the same. The highly efficient cytosine conversion is proven by the low CT values obtained when amplifying converted DNA using real-time PCR (Fig. 2). This perfect conversion rate is superior to other bisulfite kits available on the market and provides repeatable and dependable downstream analysis. Powerful DNA ProtectionDNA protection reagents are added into the DNA conversion mix solution to prevent DNA from chemical and thermophilic degradation in the bisulfite treatment and provide effective DNA denaturation, resulting in single-stranded DNA necessary for complete cytosine conversion. The prevention of DNA degradation enables subsequent amplification and analysis of large PCR fragments (Fig. 3). The efficient integrated DNA cleanup and unique elution buffer allow for long-term storage (> 6 months) and multiple freezing/thawing of the converted DNA without affecting the DNA quality.