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Published online 1 April 2004

Nucleic Acids Research, 2004, Vol. 32, No. 6 e54
© 2004 Oxford University Press

Molecular analysis of high-copy insertion sites in maize

A. Mark Settles*, Susan Latshaw and Donald R. McCarty

University of Florida, PO Box 110690, Gainesville, FL 32611–0690, USA

*To whom correspondence should be addressed. Tel: +1 352 392 7571; Fax: +1 352 392 5653; Email: amsettles{at}mail.ifas.ufl.edu
+BH417702–BH418377, CC050461–CC050722 and CG785063–CG786032

Received January 22, 2004; Revised and Accepted March 3, 2004

High-copy transposon mutagenesis is an effective tool for creating gene disruptions in maize. In order to molecularly define transposon-induced disruptions on a genome-wide scale, we optimized TAIL-PCR to amplify genomic DNA flanking maize Robertson’s Mutator insertions. Sample sequencing from 43 Mutator stocks and the W22 inbred line identified 676 non-redundant insertions, and only a small fraction of the flanking sequences showed significant similarity to maize repetitive sequences. We further designed and tested 79 arbitrary primers to identify 12 primers that amplify all Mutator insertions within a DNA sample at 3.1-fold redundancy. Importantly, the products are of sufficient size to use as substrates or probes for hybridization-based identification of gene disruptions. Our adaptation simplifies previously published TAIL-PCR protocols and should be transferable to other high-copy insertional mutagens.


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