Screen Status: Primary
Screen Overview: Primary Screen
Screener: Katharina Deiss (Parker Lab).
Extension:3505, Rm 203
Library: Full Genome 384
Library Description: Full Dharmacon 267 plate genome library stamped into 384 well plates
Library Supplier: Dharmacon
Library Format: 67 plate(s) in 384 well format
Replicate Info: 3 replicates
Publications: A genome-wide RNAi screen identifies the SMC5/6 complex as a non-redundant regulator of a Topo2a-dependent G2 arrest.. Deiss et al., 2019
Keywords: G2
Cell Line: RPE1

Screen Method

Genome Screen Protocol

Day 1: Transfection

RPE1 cells were reverse transfected with siRNA (Dharmacon) using Lullaby transfection reagent in 384 well plates.

Day 3: Treatment with 3µM ICRF193, 1µM Nocodazole

Media containing ICRF and Nocodazole compounds were added on top of the culture media.

Day 4: Cell fixation

Plates were fixed with Ethanol/Acidic (9.5:0.5) and stored at -20C.

Day 5: Staining

Cells were stained with MPM2-Cy5 and Pacific Green antibodies.

Readout

Plates are read on ArrayScan microscope and the following measurements are taken:

Plates read on Acumen Explorer and following measurements taken: