Targeted Sequencing for DNA Methylation Analysis
Zymo Research makes epigenetic biomarker validation simple with our MethylCheck™ platform. Whether you have methylation
array (27K/450K/850K) data that you would like to validate in a large sample cohort or a specific gene region in mind,
our scientists are available to design, validate, and evaluate site-specific DNA methylation changes. Simply send us your
samples and regions of interest, and we will perform every step through data analysis, sending you back publication-quality
graphs and figures.
The Targeted Bisulfite Sequencing Service Includes:
- Primer Design and Validation
- Targeted Amplification
- Adapterization and Barcoding
- Sequencing with Illumina™ Technology
- Sequence Alignment to Reference Genome
- DNA Methylation Analysis
Wilmot, et al. used the Targeted Bisulfite Sequencing service from Zymo Research to validate differentially methylated regions in childhood ADHD. The researchers extracted DNA from saliva in order to analyze methylation patterns in a sample source peripheral to brain tissue. Following genome-wide differentially methylated region analysis using bead arrays, study authors identified VIPR2 as a gene of interest for ADHD. Targeted Bisulfite Sequencing confirmed the hypomethylation of 43 sites in ADHD subjects, including VIPR2, thus providing further confidence in the use of peripheral tissue as a potential sample source for methylation analysis in psychiatric disorders.
Researchers investigating the mechanisms behind preeclampsia used the Targeted Bisulfite Sequencing service from Zymo Research to examine the DNA methylation status of three genes encoding for human galectins. Galectins found in a gene cluster on chromosome 19 are suspected to confer maternal immune tolerance to the fetus. Amplicons were designed to cover the highly methylated intragenic regions of the placental galectin genes LGALS13, LGALS14 and LGALS16. Using this strategy, several key CpG sites were found to be differentially methylated in association with preeclampsia. In conjunction with other assays, the findings suggest that differential methylation at these sites may impact or interfere with trophoblastic transcription of the cluster galectin genes.
Researchers utilized Zymo Research’s Methyl MiniSeq and Targeted Bisulfite Sequencing services to identify and validate methylation markers linked to colorectal cancer (CRC) in African Americans. Genome-wide methylation analysis with Methyl MiniSeq revealed 355 differentially methylated CpG sites in 13 genes in addition to hypomethylated Long INterspersed Elements (LINEs) in CRC samples. Furthermore, six of these genes contained the 50 CpGs with the highest degrees of differential methylation and were then studied further using 42 samples and Targeted Bisulfite Sequencing. With the larger sample set and more focused sequencing, the authors were able to validate four genes, EID3, BMP3, GAS7, and GPR75, as novel CRC methylation markers in African American patients.