Epigenetic modifications are an important pathway to better understand the association between diseases and modifications to the genome. A major epigenetic modification is DNA methylation which is known to play an important role in different biological processes. Targeted bisulfite sequencing overcomes the time-consuming and costly nature of whole genome bisulfite sequencing where 65% of the reads do not contain any CpGs.
Advantages of targeted bisulfite sequencing
- It allows researchers to specifically focus on the precise region of interest
- A targeted approach increases accuracy and sensitivity while reducing overall sequencing cost
- Using the Nimblegen capture method we ensure high uniformity and therefore optimal sequencing and sample capacity
- The product is designed to capture both strands which allows for better determination of SNPs vs. methylation status events
- In our method the bisulfite conversion is performed prior to the capture step which conserves the molecular complexity of your epigenome allowing eg for the detection of strand-specific methylation
- Availability of both fixed content (epigenome-wide design – targeting more than 5,5 million CpGs) as well as a custom design; to equally support broad discovery and focused research applications
- The customized design can be applied to any organism and regions up to 210 Mb
Targeted bisulfite sequencing workflow
First, we will perform a quality control step in which we measure the quantity and quality of the DNA provided to us by you. Only when the provided genomic DNA is of sufficient quality and quantity will we proceed with the next steps. In case the DNA provided to us is of insufficient quality and/or quantity, we will contact the customer and discuss how to proceed. We require 1µg of gDNA as input for the library preparation.
Second, we perform a restriction enzyme digest of the genomic DNA samples followed by bisulfite treatment. Afterwards a size selection of the resulting fragments is performed, followed by library preparation for sequencing. The sequencing libraries are again quality checked.
Finally, the sequencing is performed using one of our Illumina sequencers. The technical quality of the sequencing run is monitored in real time.
Sequencing data can be transferred to you via the Illumina BaseSpace platform or our own server (FTP download).
If you have more questions, please contact us.