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Originally published as Genetics Published Articles Ahead of Print on July 29, 2007.
Genetics, Vol. 177, 1163-1171, October 2007, Copyright © 2007
doi:10.1534/genetics.107.078147
High-Density Detection of Restriction-Site-Associated DNA Markers for Rapid Mapping of Mutated Loci in Neurospora
Zachary A. Lewis, Anthony L. Shiver, Nicholas Stiffler, Michael R. Miller, Eric A. Johnson and Eric U. Selker1
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229
1 Corresponding author: Institute of Molecular Biology, 1370 Franklin Blvd., Eugene, OR 97403-1229.
E-mail: selker{at}molbio.uoregon.edu
The wealth of sequence information available for Neurospora crassa and other fungi has greatly facilitated evolutionary and molecular analyses of this group. Although "reverse" genetics, in which genes are first identified by their sequence rather than by their mutant phenotypes, serves as a valuable new approach for elucidating biological processes, classical "forward" genetic analysis is still extremely useful. Unfortunately, mapping mutations and identifying the corresponding genes has typically been slow and laborious. To facilitate forward genetics in Neurospora, we have adapted microarray-based restriction-site-associated DNA (RAD) mapping for use with N. crassa oligonucleotide microarrays. This technique was used to simultaneously detect an unprecedented number of genomewide restriction site polymorphisms from two N. crassa strains: Mauriceville and Oak Ridge. Furthermore, RAD mapping was used to quickly map a previously unknown gene, defective in methylation-7 (dim-7).
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