Whole-genome sequencing and mutational analysis of human cord-blood derived stem and progenitor cells

STAR Protocols

Ruben van Boxtel¹  Eline J.M. Bertrums²  Jurrian K. de Kanter²  Anaïs J.C. N. van Leeuwen²  Axel Rosendahl Huber²  Flavia Peci² 

[1] Department of Pediatric Oncology, Erasmus Medical Center, 3015 GD Rotterdam, the Netherlands;Princess Máxima Center for Pediatric Oncology and Oncode Institute, Heidelberglaan 25, 3584 CS Utrecht, the Netherlands;

关键词: Bioinformatics;    Sequence analysis;    Cell isolation;    Single Cell;    Flow Cytometry/Mass Cytometry;    Cancer;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: Mutational signatures have been identified in cancer genomes, providing information about the causes of cancer and treatment vulnerabilities. This protocol describes an assay to determine the genotoxic mechanisms underlying these signatures using cord-blood derived hematopoietic stem and progenitor cells (CB-HSPCs). CB-HSPCs have a low mutation background, enabling sensitive detection of mutations. First, CB-HSPCs are exposed in vitro, sorted, and clonally expanded. This expansion enables whole-genome sequencing to detect the mutation load and respective patterns induced during genotoxic exposure.For complete details on the use and execution of this protocol, please refer to de Kanter et al. (2021).

【 授权许可】

Unknown