Submitted to: Crop Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/3/2012
Publication Date: 5/20/2012
Citation: Brown, R.H., Dahleen, L.S., Bregitzer, P.P. 2012. An efficient method for flanking sequence isolation in barley. Crop Science. 52:1229-1234. Interpretive Summary: One way to study what genes do in a plant is to observe what happens when the gene is made nonfunctional in a mutant. There are methods for disrupting genes with mutagenic sequences that tag the gene and thereby make it easier to figure out what gene has been disrupted. This paper presents a modified method that makes it very easy to determine which gene has been disrupted by the mutagenic sequence
Technical Abstract: An adapter ligation method was developed to determine native barley (Hordeum vulgare) sequences flanking Ds insertions and barley ESTs. This method is simple and efficient, with the majority of queries returning valid sequence information. This report describes the protocol in detail, quantifies its efficiency to enable comparisons to future modifications, and discusses specific problems that may be addressed to enable future improvements to this method.