Related Experiment Video
Updated: Feb 3, 2026

Amplicon Sequencing using the Long-Read Sequencing Technologies
Published on: August 29, 2025
Unbiased Fitness Estimation of Pooled Barcode or Amplicon Sequencing Studies
Fangfei Li1, Marc L Salit2, Sasha F Levy3
1Department of Applied Mathematics and Statistics, Stony Brook University, Stony Brook, NY 11794, USA; Laufer Center for Physical and Quantitative Biology, Stony Brook University, Stony Brook, NY 11794, USA.
Abstract:
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.
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