From: Kevin Cook <kercook@XXXX> Date: 15 June 2009 13:13:49 BST To: FlyBase-Cambridge <flybase-cambridgeXXXX>, Kim Cook <ruacookXXXX>, Stacey Christensen <sjchrist@XXXX> Subject: Isolation and characterization of Df(1)BSC824 Isolation and characterization of Df(1)BSC824 Stacey Christensen, Kim Cook and Kevin Cook Bloomington Stock Center Indiana University Df(1)BSC824 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}CG17600e01638 and P{XP}d06051. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w1118 PBac{RB}CG17600e01638/ w1118 P{XP}d06051; MKRS, P{hsFLP}86E/+ females crossed to Binsinscy/Y males. These females 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}BSC824 from the segment of PBac{RB}CG17600e01638 to the left of its FRT site and the segment of P{XP}d06051 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. The breakpoints of Df(1)BSC824 predicted from the Release 5 genomic coordinates of the transposable element insertion sites are X:21941920 ;22381239 and the cytological breakpoints predicted from these coordinates are 20C1;20F3. It failed to complement fogRA67. -- Kevin Cook, Ph.D Bloomington Drosophila Stock Center Department of Biology Indiana University 1001 E. Third St. Bloomington, IN 47405-7005 kercook@XXXX 812-856-1213 (office), 812-855-2577 (fax) http://flystocks.bio.indiana.edu
