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The colony formation assay is the gold-standard technique to assess cell viability after treatment with cytotoxic reagents, ionizing radiation, and cytotoxic combinatorial treatments. This protocol describes a high-throughput automated and high-content imaging approach to screen siRNA molecular libraries in HeLa cervical cancer cells in 96-well format. We detail reverse transfection of cells with siRNAs, followed by ionizing radiation, fixing, and staining of the plates for automated colony counting. This protocol can be used across a broad range of cell types. For complete details on the use and execution of this protocol, please refer to Tiwana et al. (2015).

Original publication

DOI

10.1016/j.xpro.2022.101355

Type

Journal article

Journal

STAR Protoc

Publication Date

17/06/2022

Volume

3

Keywords

Cancer, Cell Biology, Cell culture, Cell-based Assays, High Throughput Screening, Molecular Biology, Single Cell