The need for advanced DNA methylation analysis in translational research and clinical settings is increasingly critical, especially in oncology. Liquid biopsies, with their non-invasive nature, offer a promising avenue for cancer diagnostics and monitoring. However, the fragmented DNA in these samples poses analytical challenges.
By learning how to detect fully methylated, unmethylated, and mixed methylation sites, we can gain crucial insights into genomic heterozygosity, epigenetic variation, and tumor heterogeneity. These insights are vital for understanding complex biological processes and disease mechanisms, directly impacting patient care and treatment strategies.
Sanger sequencing provides an attractive option for analyzing methylated DNA sequences. The workflow is relatively simple, fast, and low cost. Data analysis of the peaks generated in an electropherogram is relatively straightforward; a reference genome based on bisulfite-modified DNA is not needed. Additionally, a single reaction can give information about all positions in the amplicon, facilitating the detection of partially modified sequences.
Through Sanger sequencing, translational research and clinical labs can play a pivotal role in improving patient outcomes and advancing cancer research.
Download the full white paper to view the full method for analyzing DNA methylation in liquid biopsy samples using Sanger sequencing.
A protocol is also provided in the appendix that can be used to bisulfite convert input DNA, amplify specific loci, and sequence regions of interest to detect changes in methylation.
