Reduced Representation Bisulfite Sequencing – RRBS

Reduced representation bisulfite sequencing (RRBS) is an efficient and high-throughput technique used to analyze the genome-wide methylation profiles on a single nucleotide level. This technique combines a methylation independent restriction enzyme digest and bisulfite sequencing  in order to enrich for the areas of the genome that have a high CpG content. The fragments that comprise the reduced genome include the majority of promotors, as well as regions such as repeat sequences that are difficult to profile using conventional bisulfite sequencing approaches.

Our RRBS service provides our customers with the single-base resolution methylation profile throughout the genome (the Methylome) of their samples.

Highlights of this service:

–          Genome-wide

–          True single-base resolution

–          Applicable to all mammalian samples and tissue types

–          Cost-effective technique

The Workflow:

The process consists of several steps.

We start the process with performing a first quality control step in which we measure the quantity and quality of the DNA provided to us by our customers. 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.

In a second step we perform a  restriction enzyme digest of the genomic DNA samples followed by library preparation for sequencing. Afterwards a size selection of the resulting fragments is performed.

After the size selection bisulfite conversion is executed.

The sequencing is then performed using Illumina sequencing technology.

Upon completion of the sequencing run the bio-informatics part of the service is started by performing basecalling, cleaning and mapping of the data. Afterwards the data is uploaded into our Genome Browser software. Customers get access to the data files for their samples together with access codes to the Genome Browser software in which they can analyze their data in an intuitive and easy-to-use software environment.