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Genome Res. 2009 Oct;19(10):1836-42. doi: 10.1101/gr.093955.109. Epub 2009 Jul 21.

Quantitative phenotyping via deep barcode sequencing.

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  • 1Department of Molecular Genetics, University of Toronto, Ontario, Canada.

Abstract

Next-generation DNA sequencing technologies have revolutionized diverse genomics applications, including de novo genome sequencing, SNP detection, chromatin immunoprecipitation, and transcriptome analysis. Here we apply deep sequencing to genome-scale fitness profiling to evaluate yeast strain collections in parallel. This method, Barcode analysis by Sequencing, or "Bar-seq," outperforms the current benchmark barcode microarray assay in terms of both dynamic range and throughput. When applied to a complex chemogenomic assay, Bar-seq quantitatively identifies drug targets, with performance superior to the benchmark microarray assay. We also show that Bar-seq is well-suited for a multiplex format. We completely re-sequenced and re-annotated the yeast deletion collection using deep sequencing, found that approximately 20% of the barcodes and common priming sequences varied from expectation, and used this revised list of barcode sequences to improve data quality. Together, this new assay and analysis routine provide a deep-sequencing-based toolkit for identifying gene-environment interactions on a genome-wide scale.

PMID:
19622793
[PubMed - indexed for MEDLINE]
PMCID:
PMC2765281
Free PMC Article

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