In conjuction with the Cells-to-CpG™ Bisulfite Conversion Kit, use the Cells-to-CpG™ Methylated and Unmethylated gDNA Control Kit to prepare methylated DNA standards with known levels of methylation. Perform bisulfite conversion of the methylated DNA standards, then use the converted methylated DNA standards when you perform downstream analysis to determine the percentage of methylated DNA in your samples.
The bisulfite method is the most commonly used technique for identifying specific methylation patterns within a DNA sample. It consists of treating DNA with bisulfite, which converts unmethylated cytosines to uracil but does not change methylated cytosines (Figure 1). Bisulfite conversion has been utilized in DNA methylation research for the last 20 years with few improvements to the technology until now. With thorough optimization, the Cells-to-CpG™ Bisulfite Conversion Kit provides a quick, streamlined method (Figure 2) for bisulfite conversion to reveal methylated cytosines in either loci-specific or genomewide analyses.
Key product features:
• Flexible DNA conversion—validated solution for Life Technologies platforms to minimize the need for optimization
• Quantitative reliability—confidence in results with available controls to easily monitor conversion rate
• Streamlined workflow—ability to start with various sample input types including cell, tissue, blood, and FFPE samples
• Efficient application—reduced time and labor through direct conversion of cytosines in samples without need for purifying genomic DNA
Complete DNA Conversion with Greater Flexibility
By including all of the components from direct sample conversion to purification, the Cells-to-CpG™ System provides a validated, flexible, simplified bisulfite conversion solution that is ideal for real-time PCR, CE, and next generation sequencing workflows. Choose a 50-reaction format or 2x96-plate format for more flexibility in performing your bisulfite conversion. Validated controls, including the Cells-to-CpG™ Bisulfite Conversion and Quantitation Control Kit as well as the Cells-to-CpG™ Methylated and Unmethylated Control Kit, are available to use through bisulfite conversion and amplification of the bisulfite-converted DNA.
Quantitatively Reliable Results
The Cells-To-CpG™ Bisulfite Conversion Kit is designed to provide quantitatively reliable results verified with available controls. DNA recovery after conversion correlates linearly to the input amount with no bias observed between non-methylated and methylated samples (Figure 3).
Streamlined Bisulfite Conversion Workflow with Efficient Application
Starting with cells, tissues, blood, FFPE samples, or purified DNA, directly lyse and denature sample for subsequent bisulfite conversion, desalting, and desulfonation. The protocol takes less than 3 hours, which is a significant reduction in time compared to other available methods that require at least 18 hours.