Methylated sequence capturing using a Methyl Binding Domain (MBD) based enrichment followed by next-generation sequencing provides a great combination of sensitivity and cost-efficiency for genome-wide DNA-methylation profiling. The use of methyl-CpG binding domain (MBD) proteins for affinity based purification is believed to be an inherently good technique due to the biological origin of the MBD. On top of this, recent studies have shown that MBD-seq (also called MethylCap-seq or MiGS) is great all-round technique for genome-wide DNA methylation studies.
Our “MBD-based genome-wide methylation sequencing” service provides our customers with the methylation profile throughout the entire genome (the Methylome) of their samples. The resulting data allows our customers to screen for “Differentially Methylated Regions” in a genome-wide approach for the specific disorder, event or phenotype they are studying.
Highlights of this service:
– True genome-wide
– Approximate basepair resolution
– 100 times more sensitive than MeDIP
– >95% of genome-wide BSS data is also found with this service
– Low DNA input (500ng genomic DNA)
– Over 3000 methylomes sequenced
– Applicable to all mammalian samples and tissue types
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 random fragmentation of the genomic DNA samples followed by size selection of the resulting fragments. This is followed by a second quality control step in which the resulting DNA concentration is determined and the DNA is checked for degradation.
The resulting DNA fragments will then be separated by performing MBD-capture during which all fragments containing methylation are withheld, while fragments that do not contain methylation are discarded. Once again the DNA concentration will be determined during as a quality control step.
The library preparation and sequencing are 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.