|Rinehart, Timothy - Tim|
Submitted to: Annual International Plant & Animal Genome Conference
Publication Type: Abstract Only
Publication Acceptance Date: 10/5/2005
Publication Date: 1/17/2006
Citation: Wang, X., Trigiano, R.N., Windham, M.T., Devries, R., Rinehart, T.A., Spiers, J.M., Scheffler, B.E. 2006. The optimization of microsatellite discovery in flowering dogwood (cornus florida l.). Annual International Plant & Animal Genome Conference.
Technical Abstract: Microsatellite loci are useful genetic markers for studying genetic diversity and for creating linkage maps of flowering dogwood. Microsatellite discovery is tedious work and expensive. We modified existing techniques and developed an efficient method for identifying many microsatellite loci in flowering dogwood. The modifications include rapid construction of a small insert enriched-microsatellite library and a simple procedure for microsatellite discovery without colony hybridization. Genomic DNA was digested with 1-3 restricted enzymes to produce 300-800 bp fragments. Without any end modification, fragments were ligated with an artificial double-strand adapter. The fragments containing microsatellite motifs were enriched with streptavidin-coasted magnetic beads and cloned into a pBluescript II (+) vector to construct an enriched-microsatellite library. To screen for clones containing microsatellites from the enriched library, a simple PCR amplification with a combination of vector universal primers and a synthesized microsatellite oligo was performed. The colonies in which PCR