Unbiased Fitness Estimation of Pooled Barcode or Amplicon Sequencing Studies.

Standard practice for phenotyping complex cell pools is to measure the fold enrichment of genotype-specific amplicons after a period of competitive growth. Here, we show that fold-enrichment measures cannot be compared across genotype pools with different fitness distributions. We develop a method to calculate an unbiased estimate of relative fitness by tracking abundances over several time points and show how to optimize experimental protocols to minimize fitness measurement error.