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Methods Mol Biol. 2012;910:55-69. doi: 10.1007/978-1-61779-965-5_4.

Barcode sequencing for understanding drug-gene interactions.

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  • 1Donnelly Centre, University of Toronto, Toronto, ON, Canada.

Abstract

With the advent of next-generation sequencing (NGS) technology, methods previously developed for microarrays have been adapted for use by NGS. Here we describe in detail a protocol for Barcode analysis by sequencing (Bar-seq) to assess pooled competitive growth of individually barcoded yeast deletion mutants. This protocol has been optimized on two sequencing platforms: Illumina's Genome Analyzer IIx/HiSeq2000 and Life Technologies SOLiD3/5500. In addition, we provide guidelines for assessment of human knockdown cells using short-hairpin RNAs (shRNA) and an Illumina sequencing readout.

PMID:
22821592
[PubMed - indexed for MEDLINE]
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