AIM-Seq: A simple method to rapidly map random insertional mutations
The Cryptococcus neoformans strain H99 was randomly mutagenized by Agrobacterium tumefaciens-mediated transformation  with  Agrobacterium tumefaciens strain EHA105 expressing the PZP-NATcc plasmid.  The mutant collection was screened for sensitivity to high pH or high salt, or resistance to sodium dodecyl sulfate, phenotypes indicating cell wall defect.  27 mutants with one or more of these phenotypes were selected.  DNA was extracted from each, then pooled in equimolar amounts for library preparation and sequencing by Illumina MiSeq.