title: Scaling down: a PCR-based method to efficiently screen for desired knockouts in a high density Mycobacterium tuberculosis picked mutant library.
abstract: Transposon mutagenesis produces random mycobacterial mutants at high frequency. Because they are random, however, it is difficult to isolate mutations in particular target genes. Here we describe the use of an arrayed library of Mycobacterium tuberculosis together with a PCR screening strategy to rapidly identify strains with defined insertion mutations. This method is useful for many genetic applications.
authors.affiliation: Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: James M
authors.initials: JM
authors.lastname: Lane
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Rubin
mesh_terms: D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D015723:Gene Library; D005798:Genes, Bacterial; D016254:Mutagenesis, Insertional; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial