Thursday, February 20th, 2014
David Gresham has led a collaboration with John Storey at Princeton University and Wei Chen at the Max Delbruck Center to develop optimized experimental and computational methods for studying complex mixtures of mutants using next-generation sequencing. BAR-seq entails sequencing molecular barcodes that uniquely identify engineered yeast mutants. By analyzing a mixture of 4,500 knock out mutants using BAR-seq and simulating different experimental designs in silico the researchers discovered that a surprisingly low sequence read depth provides sufficient statistical power to analyze 30 genome-wide screens in a single sequencing lane. The findings are relevant for study designs using other methods for pooled analysis of mutants including Tn-seq and large-scale mutagenesis using (CRISPR)-Cas9. The study was published in the open access journal of the Genetics Society of America, G3.
