Automated 96-well format high throughput colony formation assay for siRNA library screen

被引：0

作者：

Hatch, Stephanie B. ^{[1
]}

Prevo, Remko ^{[2
]}

Chan, Tiffany ^{[2
]}

Millar, Val ^{[1
]}

Cornelissen, Bart ^{[2
]}

Higgins, Geoff ^{[2
]}

Ebner, Daniel ^{[1
]}

机构：

[1] Univ Oxford, Target Discovery Inst, Nuffield Dept Med, Oxford, England

[2] Univ Oxford, Dept Biol, Oxford, England

来源：

STAR PROTOCOLS | 2022年 / 3卷 / 02期

基金：

英国医学研究理事会;

关键词：

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

D O I：

10.1016/j.xpro.2022.101355

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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).

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页数：16

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