BioSample

Transposition events of an endogenous, single-copy Dissociation (Ds) element were selected by a two step process. First, kernels were scored for Ds excision from a donor locus. Then to detect Ds reinsertion, genomic DNA was extracted from F2 segregants that lacked Ac and was analyzed by gel blots with methylation sensitive restriction enzymes and a Ds probe. DNA flanking one side of the transposed Ds was recovered by IPCR, gel-purified and sequenced directly with nested primers from one or both sides of the fragment. A few fragments required additional PCR or cloning steps to generate sequence. The obtained sequence was processed to remove low quality, vector and Ds-derived sequences, to assemble reads into contigs, and to correlate the sequence with information derived from the Southern blot and IPCR steps. For additional details see the project web page at http://plantgdb.org/prj/AcDsTagging/index.php.