Signature-tagged mutagenesis (1995)

Hensel M, Shea JE, Gleeson C, Jones MD, Dalton E, Holden DW. Simultaneous identification of bacterial virulence genes by negative selection. Science. 1995 Jul 21;269(5222):400-3. PubMed PMID: 7618105.

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As a result of the ability to assay multiple mutants in a multiplex fashion, the technique was widely adopted (reviewed by Joan Mecsas). However, there are a number of technical obstacles that limited the utility of the technique: - Assay of at most 96 mutants in each assay (limited by the hybridization approach). - Tedious library construction: each well is derived from a different transposon. - Not quantitative. - Radiolabeling is required for detection.

So on to multiplex hybridization…!

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Next: Microarray Hybridization (2001)