GSM4120372: DMCPD30mR1; Drosophila melanogaster; OTHER
S2-DGRC cells were obtained from the Drosophila Genetic Resources Center. Cells were cultured in Schneider's medium at 27 degrees with heat inactivated FBS at 10% (v/v).  For UV treatments, cells were inoculated into R-150 plates and grown to about 25 to 80 percent confluence.  Media was gently removed from the semi-adherent monolayer, cells were irradiated with UV-C, and then fresh media containing sterilized, conditioned media was added to cells, and cells were incubated at 27 degrees.  XR-seq assays employed 20 Jm-2.    Following predetermined repair times, plates were placed on ice and cooled cells were harvested by scraping and rinsing with ice-cold PBS. Cells were pelleted, transferred to Eppendorf tubes, washed with cold PBS, and resuspended in 320 to 340 ul of cold TE.  Samples were processed using 5 ul RNase A and proteinase K. Samples were immunoprecipitated with anti-CPD antibody, and then ligated to adapters and processed for sequencing (XR-seq).  One plate of S2 cells per repair time point was sufficient for XR-seq assay.  Drosophila possesses photolyases, so cells were kept in the dark or under dim yellow illumination from the time of irradiation to the time of addition of NaCl to cell lysates. After PCR amplification, libraries wrere sequencing on HS4000.
GEO Accession
GSM4120372