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• W4283717526 abstract "Arrayed libraries of defined mutants have been used to elucidate gene function in the post-genomic era. Yeast haploid gene deletion libraries have pioneered this effort, but are costly to construct, do not reveal phenotypes that may occur with partial gene function and lack essential genes required for growth. We therefore devised an efficient method to construct a library of barcoded insertion mutants with a wider range of phenotypes that can be generalized to other organisms or collections of DNA samples. We developed a novel but simple three-dimensional pooling and multiplexed sequencing approach that leveraged sequence information to reduce the number of required sequencing reactions by orders of magnitude, and were able to identify the barcode sequences and DNA insertion sites of 4391 Schizosaccharomyces pombe insertion mutations with only 40 sequencing preparations. The insertion mutations are in the genes and untranslated regions of nonessential, essential and noncoding RNA genes, and produced a wider range of phenotypes compared to the cognate deletion mutants, including novel phenotypes. This mutant library represents both a proof of principle for an efficient method to produce novel mutant libraries and a valuable resource for the S. pombe research community." @default.
• W4283717526 created "2022-07-01" @default.
• W4283717526 creator A5027118136 @default.
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• W4283717526 date "2022-06-29" @default.
• W4283717526 modified "2023-10-15" @default.
• W4283717526 title "A multiplexed, three-dimensional pooling and next-generation sequencing strategy for creating barcoded mutant arrays: construction of a <i>Schizosaccharomyces pombe</i> transposon insertion library" @default.
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• W4283717526 doi "https://doi.org/10.1093/nar/gkac546" @default.
• W4283717526 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/35766443" @default.
• W4283717526 hasPublicationYear "2022" @default.
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