From: Kevin Cook <kercook@XXXX> Date: 6 May 2009 00:43:19 BST To: flybase-cambridgeXXXX, sjchristXXXX, ruacook@XXXX Subject: Isolation and characterization of Df(3L)BSC796 Isolation and characterization of Df(3L)BSC796 Stacey Christensen, Kim Cook and Kevin Cook Bloomington Stock Center Indiana University Df(3L)BSC796 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}mRpL15e03567 and P{XP}d06749. 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; PBac{RB}mRpL15e03567/P{XP}d06749 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}BSC796 from the segment of PBac{RB}mRpL15e03567 to the left of its FRT site and the segment of P{XP}d06749 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(3L)BSC796 predicted from the Release 5 genomic coordinates of the transposable element insertion sites are 3L:20429215 ;20723658 and the cytological breakpoints predicted from these coordinates are 77C3;77E4. Df(3L)BSC796 failed to complement kniri-1 and Df(3L)BSC448. -- 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