From: Kevin Cook <kcook@XXXX> To: FlyBase-Cambridge <flybase-cambridgeXXXX>, kaufmanXXXX, Stacey Christensen <sjchristXXXX>, Kim Cook <ruacookXXXX> Subject: Isolation and characterization of Df(1)BSC641 Date: Mon, 20 Oct 2008 09:19:36 -0400 ( 14:19 BST) Isolation and characterization of Df(1)BSC641 Kim Cook, Stacey Christensen and Kevin Cook Bloomington Stock Center Indiana University Df(1)BSC641 was isolated as a FLP recombinase-induced recombination event involving P{XP}Fimd03334 and PBac{RB}CG8142e04583. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{XP}Fimd03334/PBac{RB}CG8142e04583; 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}BSC641 from the segment of P{XP}Fimd03334 to the left of its FRT site and the segment of PBac{RB}CG8142e04583 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 cytological breakpoints of Df(1)BSC641 predicted from the Release 5 genomic coordinates of the transposable element insertions sites are 15F9;16E1. The presence of a deletion was confirmed cytologically, though the breakpoints were not analyzed in detail. __________________________________________________________ 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
