From: kcook@XXXX Subject: Isolation and characterization of Df(3R)BSC682 Date: December 28, 2008 1:15:51 PM GMT- 05:00 Isolation and characterization of Df(3R)BSC682 Stacey Christensen, Kim Cook and Kevin Cook Bloomington Stock Center Indiana University Df(3R)BSC682 was isolated as a FLP recombinase-induced recombination event involving P{XP}d00236 and PBac{RB}e01013. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w1118; P{hs-hid}3, Dr1/TM6C, Sb1 cu1 females crossed to P{hsFLP}1, y1 w1118; P{XP}d00236/PBac{RB}e01013 males. The males were heat shocked as larvae as described in Parks et al., Nature Genetics 36: 288-292, 2004 (FBrf0175003). This cross and crosses in preceding and succeeding generations maintained the original genetic background of the Exelixis insertion stocks (Thibault et al., Nature Genetics 36: 283-287, 2004; FBrf0175002). The recombination event generated the genetic element P+PBac{XP5.RB3}BSC682 from the segment of P{XP}d00236 to the left of its FRT site and the segment of PBac{RB}e01013 to the right of its FRT site. Its presence was verified using the PCR methods and primers described in Parks et al. with the substitution of the primer 5'-GCTTCTAAACGCTTACGCATAAACGATG-3' for the RB3' plus or RB3' minus primer in the Hybrid PCR protocol in the Supplementary Methods. Exelixis, Inc. determined the insertion site of P{XP}d00236 to be Release 3 genomic coordinate 14022167 on chromosome arm 3R. This corresponds to 90F6 on the Release 5 genome map. The predicted position of PBac{RB}e01013 on the Release 5 map is 91A1. Consequently, the cytological breakpoints of Df(3R)BSC682 are predicted to be 90F1;91A1. Df(3R)BSC682 failed to complement repo03702. __________________________________________________________ Kevin Cook, Ph.D. Bloomington Drosophila Stock Center Department of Biology http://flystocks.bio.indiana.edu Jordan Hall 142 Indiana University 812-856-1213 1001 E. Third St. 812-855-2577 (fax) Bloomington, IN 47405-7005 kcook@XXXX