Dvořák Tomaštíková, E., Pecinka, A.
Methods for plant nucleus and chromatin studies : 53-70, 2025

Keywords: Genetic mapping, Forward genetics, Mapping-by-sequencing, High-throughput sequencing, SNP calling, DNA damage repair, Zebularine, DNA-protein crosslinks
Abstract: Forward-directed genetic screens are extremely powerful in identifying novel genes involved in a specific biological process, including various chromatin regulatory pathways. However, the traditional ways of genetic mapping are time- and cost-demanding. Recently, the whole process was revolutionized by the development of mapping-by-sequencing (MBS) protocols. In MBS, the causal mutations and their positions within genes are identified directly by whole-genome sequencing and bioinformatics analysis of the bulk of mutant plants selected based on the mutant phenotype from a segregating population. MBS increases precision and economizes the mapping. Here, we describe a general protocol and provide practical tips on how to proceed with the mapping-by-sequencing on the example of Arabidopsis forward-directed genetic screen designed to identify mutants sensitive to a specific type of DNA damage. The described protocol is generally applicable to a wide range of genetic screens in various inbreeding species with a reference genome sequence.
DOI: 10.1007/978-1-0716-4228-3_4
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IEB authors: Aleš Pečinka, Eva Dvořák Tomašt...