Highly replicated experiments studying complex genotypes using nested DNA barcodes.

ABSTRACT: Many biological experiments involve studying the differences caused by genetic modifications, including genotypes composed of modifications at more than one locus. However, as the number and complexity of the genotypes increases, independently generating and tracking the necessary number of biological replicate samples becomes a major challenge. We developed a barcode-based method to track large numbers of independent replicates of combinatorial genotypes in a pooled format, enabling robust detection of subtle phenotypic differences. To construct a plasmid library of combinatorial genotypes, we utilized a nested serial cloning process to combine gene variants of interest that have associated DNA barcodes. The final plasmids each contain variants of multiple genes of interest, and a combine

SUBMITTER: Monge M 

PROVIDER: S-EPMC11956976 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature