December 15, 2017

(Oxidative) Reduced Representation Bisulfite Sequencing

(Oxidative) reduced representation bisulfite sequencing [(ox)RRBS] is an efficient and high-throughput technique used to analyze the genome-wide methylation profiles on a single nucleotide level. This technique combines methylation independent restriction enzymes and bisulfite sequencing in order to enrich for the areas of the genome that have a high CpG content. This reduces the amount of nucleotides needed to be sequenced to 1% of the genome. The fragments that comprise the reduced genome still include the majority of promoters, as well as regions such as repeated sequences that are difficult to profile using conventional bisulfite sequencing approaches.

Advantages of (ox)RRBS

  • True single-base resolution
  • Applicable to all mammalian samples and tissue types, also to other non-mammalian organisms that have a good reference genome
  • Cost-effective technique
  • Availability to offer support during BSS primer design and PCR optimization

(ox)RRBS 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.

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.