Competition assays using barcoded deletion strains to gain insight into small RNA function.
Ordered collections of barcoded deletion mutants can be screened rapidly in mixed cultures to uncover resistance and sensitivity phenotypes associated with the loss of a gene. As such, they are invaluable tools for assigning gene function in the post-genomic era. In this protocol, we describe methodologies for creating and employing barcoded deletion mutants in competitive screens as well as how to analyze the ensuing results.
Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD, USA.
Errett C
EC
Hobbs
Gisela
G
Storz
D004269:DNA, Bacterial / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D016680:Genome, Bacterial / Q000235:genetics; D016254:Mutagenesis, Insertional / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D012329:RNA, Bacterial / Q000235:genetics* / Q000378:metabolism*; D022661:RNA, Untranslated / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion / Q000235:genetics*; D013997:Time Factors
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D022661:RNA, Untranslated