| Screen Status: | Primary Repeat |
| Screen Overview: | Primary genome-wide screen in MAD heterozygote cell line |
| Screener: | Alvin Lee (Swanton Lab). Extension:3462, Rm 115 |
| Library: | Full Genome 96 |
| Library Description: | 270 plates consisting of Kinome, GPCR, druggable and genome libraries |
| Library Supplier: | Dharmacon |
| Library Format: | 267 plate(s) in 96 well format |
| Replicate Info: | 3 replicates |
| Publications: | Ccdc13 is a novel human centriolar satellite protein required for ciliogenesis and genome stability.. Staples et al., 2014 |
| Keywords: | CIN, MAD2 |
| Cell Line: | HCT116 |
The screen is performed in 96well format with poly-L-Lysine coated plates for better cell attachment.
Day 1: HCT116 cell transfection
HCT116 wild-type and Mad2+/- cells were transfected using lipoRNAiMAXi transfection reagent, then cultured for 4 days.
Day 4: Ethanol fixation
Plates were fixed with 80% EthOH.
Staining and Readout
Cells were stained with pH2AX antibody and DAPI.
pH2AX nuclear intensity and total cell number per well were determined using the Acumen Explorer. Also, pH2AX nuclear spots were quantified using Cellomics Arrayscan.