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PubMed · 39576596

A Practical Approach to High-Throughput and Accurate Mapping-by-Sequencing in Arabidopsis.

Eva Dvořák Tomaštíková·Ales Pecinka
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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.

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ArabidopsisChromosome MappingHigh-Throughput Nucleotide SequencingGenome, PlantComputational BiologyMutationPhenotypeWhole Genome SequencingDNA damage repairDNA-protein crosslinksForward geneticsGenetic mappingHigh-throughput sequencingMapping-by-sequencingSNP callingZebularineJournal ArticleResearch Support, Non-U.S. Gov't

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Eva Dvořák Tomaštíková, Ales Pecinka. 2025. A Practical Approach to High-Throughput and Accurate Mapping-by-Sequencing in Arabidopsis.. https://doi.org/10.1007/978-1-0716-4228-3_4

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