DNA Methylation Profiling via Whole Genome Bisulfite-Seq
Aberrant DNA methylation has been implicated in numerous human diseases including cancer and genetic disorders. Recent advance in DNA methylation technologies has enable profiling of the entire methylome at single base resolution. We provide Bisulfite-seq ready Solexa PE library construction, and whole genome bisulfite sequencing on Solexa GAII platform. We also provide informatic support. Please contact us for more information.
Bisulfite-seq PE library is made by ligating methylated PE (mPE) adaptor to fragmented genomic DNA. The mPE adaptor-ligated genomic DNA is then bisulfite treated twice, resulting in conversion rate of > 99% of unmethylated cytosine. Bisulfite converted library is then recovered by limited cycles of PCR amplification. The final library size of around 200-300 bp is gel purified and quantified via fluorometer. We QC the library quality by random subcloning and sequencing of ~20-30 colonies to check for proper library construction and bisulfite conversion. The QC-ed library is then ready for cluster formation and high throughput Illumina sequencing.
We provide custom data analysis including:
1. Alignment to the reference genome
2. Determine methylation ration for each sequenced cytosine
3. Statistical analysis of differential methylation between two samples or tissue types.
4. Visual distribution of the differentially methylated regions.
5. Other custom analyses.