JSON
Micallef P, Santamaría ML, Escobar M, Andersson D, Österlund T, Mouhanna P, Filges S, Johansson G, Fagman H, Vannas C, Ståhlberg A
Genome Biol. 26 (1) 37 [2025-02-25; online 2025-02-25]
Digital sequencing uses unique molecular identifiers (UMIs) to correct for polymerase induced errors and amplification biases. Here, we design 19 different structured UMIs to minimize the capacity of primers to form non-specific PCR products during library construction using SiMSen-Seq, a PCR-based digital sequencing approach with flexible multiplexing capabilities suitable for tumor-informed mutation analysis. All structured UMI designs demonstrate enhanced assay performance compared with an unstructured reference UMI. The best performing structured UMI design shows significant improvements in all tested aspects of assay and sequencing performance with the ability to reliable detect low variant allele frequencies.
NGI Uppsala (SNP&SEQ Technology Platform) [Service]
National Genomics Infrastructure [Service]
PubMed 40001095
DOI 10.1186/s13059-025-03504-x
Crossref 10.1186/s13059-025-03504-x
pmc: PMC11853513
pii: 10.1186/s13059-025-03504-x