Testing for de novo mutation calling artefacts by re-sequencing ten siblings of a single mutation accumulation line in Arabidopsis thaliana. Arabidopsis_mutation_accumulation_siblings

We conducted a mutation bias study in the plant Arabidopsis thaliana. To test for the possibility that our mutation bias results could in part be artefacts of a pooled-seedling sequencing approach, we resequenced entire rosettes of siblings (individual plants) of one mutation accumulation line (#73) and asked if the distribution of variation called (i.e., putative somatic mutations around transcription start and termination sites) was similar to the patterns seen with the seedling pools of the 107 individual lines described in the preceding section. Specifically, we grew 10 siblings of line #73 and extracted DNA from 3-week old whole rosettes. Barcoded PCR-free libraries for the 10 siblings were sequenced, with 150 bp paired-end reads, at approximately 60x depth each on a single lane of the Illumina HiSeq 3000 platform. Additionally, for one sibling, the same library was sequenced in an independent lane at approximately 600x depth.