From: Kevin Cook <kcook@XXXX> To: FlyBase-Cambridge <flybase-cambridgeXXXX>, Kim Cook <ruacookXXXX>, Stacey Christensen <sjchristXXXX>, kaufmanXXXX Subject: Isolation and characterization of Df(3L)BSC563 Date: Wed, 03 Sep 2008 11:58:38 -0400 ( 16:58 BST) Isolation and characterization of Df(3L)BSC563 Kim Cook, Stacey Christensen and Kevin Cook Bloomington Stock Center Indiana University Df(3L)BSC563 was isolated as a FLP recombinase-induced recombination event involving P{XP}d00582 and PBac{RB}CG11396[e01526]. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w[1118]; P{hs-hid}3, Dr[1]/TM6C,Sb[1] females crossed to P{hsFLP}1, y[1] w[1118]; P{XP}d00582/PBac{RB}CG11396[e01526] 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}BSC563 from the segment of P{XP}d00582 to the left of its FRT site and the segment of PBac{RB}CG11396[e01526] 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'-CCAATGCGTTTATTTCAGGTCACG-3' for the RB3' plus or RB3' minus primer in the Hybrid PCR protocol in the Supplementary Methods. The cytological breakpoints of Df(3L)BSC563 predicted from the Release 5 genomic coordinates of the transposable element insertions sites are 77E1;77E6. Df(3L)BSC563 failed to complement kni[ri-1]. __________________________________________________________ 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
