Highly replicated experiments studying complex genotypes using nested DNA barcodes.
We developed a barcode-based method to track large numbers of independent replicates of a small number of combinatorial genotypes in a pooled format. We utilized a nesting serial cloning process to combine gene variants of interest that have associated DNA barcodes. Using this approach, we tested the functionality of combinations of yeast, human, and null orthologs of the nucleotide excision repair factor (NEF-1) complex. Our NICR (nested identification combined with replication) barcoding method allows for high-throughput combinatorial genetic screens.