Bisulfite converted DNA generated with EpiTect Kits was stored at –20°C for up to 36 months, and amplified at several time intervals. The resulting threshold cycle values at all time points for various DNA amounts were comparable to those obtained with freshly converted DNA (time point 0).
[A] Upon addition to the bisulfite conversion reaction, the DNA Protect Buffer [B] should turn from green to blue, indicating sufficient mixing and correct pH.
DNA was purified from rat liver FFPE tissue (10 µm) using the EpiTect Plus FFPE Bisulfite Kit (1–4) or an alternative kit from Supplier Z (5–8). [A] Detection of the indicated genes using the PyroMark PCR Kit and agarose gel analysis. [B] Detection of two amplicons of the GSTP gene (156 and 279 bp) using the QuantiTect SYBR® Green PCR Kit (4 triplicate samples). Templates purified using the EpiTect Plus FFPE Bisulfite Kit resulted in lower CT values and highly reproducible results for both amplicons (left). Templates purified using the kit from Supplier Z resulted in amplification of only the 156 bp fragment with higher CT values (right).
Degradation of DNA during the bisulfite conversion reaction results in low yields of the target sequence for downstream applications. This problem is particularly important for samples such as FFPE tissues, where the starting DNA is already fragmented. The DNA Protect Buffer included in EpiTect Plus Bisulfite Kits minimizes DNA damage during conversion. Along with minimal loss of DNA during sample lysis and thorough cleanup of the resulting converted DNA, this innovative DNA protect technology maximizes recovery of the converted DNA.
Streamlined and straightforward, the EpiTect Plus DNA Bisulfite Kit enables complete bisulfite conversion and clean up of genomic DNA in approximately 6 hours.
Upon completion of the sample lysis steps, bisulfite conversion is completed using the included EpiTect Plus DNA Bisulfite Kit.
Upon completing deparaffinization, lysis, and decrosslinking steps, bisulfite conversion is completed using the EpiTect Plus DNA Bisulfite Kit.
Depending on sample type, the bisulfite conversion of DNA calls for the use of a specific kit and protocol. EpiTect Plus Bisulfite Kits contain buffers optimized for DNA extraction and all reagnets for the DNA bisulfite conversion.