Sanger sequencing method is less sensitive and maybe not suitable for the detection of mutations in ctDNA. However, In case to detect mutations in some samples (ctDNA) with sanger seq; mutant allele frequency (MAF) could be identified in this technique?
Sanger sequencing will pick up mutations in circulation dna. This dna is often quite short and the dna template is enriched by using dna purification kits which are designed to purify short or viral dna fragments. The existence of 2 overlapping peaks in the electropherogram indicates 2 species (normal and mutant) are present and if there are SNPs present in the gene close to the mutation then ARMS pcr may further enrich the signal from the tumour over that of the normal cell dna
I want to asked help of specialists in these technologies in my field of research, and I also want to refer to detailed research in 2020 that addressed this question with advanced and accurate research.
Article Sanger Validation of High-Throughput Sequencing in Genetic D...
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