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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
Published on: March 31, 2022
synapsis: A Bioconductor Package to Automate the Analysis of Meiotic Double-Strand Break Repair and Crossover
Lucy McNeill1, Vanessa Tsui1,2, Wayne Crismani3,4
1DNA Repair & Recombination Laboratory, St Vincent's Institute of Medical Research, Fitzroy, VIC, Australia.
Abstract:
Immunofluorescent staining is commonly used to generate images to characterize cytological phenotypes. The manual quantification of DNA double-strand breaks and their repair intermediates during meiosis using image data requires a series of subjective steps, from image selection to the counting of particular events per nucleus. Here we describe "synapsis," a bioconductor package, which includes a set of functions to automate the process of identifying meiotic nuclei and quantifying key double-strand break formation and repair events in a rapid, scalable, and reproducible workflow, and compare it to manual user quantification. The software can be extended for other applications in meiosis research, such as incorporating machine learning approaches to categorize meiotic substages.
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