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Identifying crossover locations in an Arabidopsis thaliana Ler x Col/Ler backcross F1 population using genotyping by sequencing for male-specific crossover map

ABSTRACT: Genomic DNA from 96 Ler x Col/Ler backcross F1 population was extracted using CTAB and used to generate sequencing libraries as described (Rowan et al; Yelina et al), with the following modifications. DNA was extracted from 3 rosette leaves of 5 week old plants and 150 ng of DNA used as input for each library. DNA shearing was carried out for 20 minutes at 37°C with 0.4U of DNA Shearase (Zymo research). The barcoded adapters used for library construction are listed in Rowan et al. Each set of 96 libraries was sequenced on one lane of an Illumina NextSeq500 instrument (300-cycle Mid Output run). Sequencing data was analysed to identify crossovers as previously reported, using the TIGER pipeline.

INSTRUMENT(S): HiSeq X Ten

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Kyuha Choi

PROVIDER: E-MTAB-12695 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Identifying crossover locations in an Arabidopsis thaliana Col/Ler x Ler backcross F1 population using genotyping by sequencing for female-specific crossover map

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Project description:Genomic DNA from 144 wild type Col x Ler F2 individual and 96 meiMIGS-PPX1-PPX2 Col x Ler F2 individuals was extracted using CTAB and used to generate sequencing libraries as described (Rowan et al; Yelina et al), with the following modifications. DNA was extracted from 3 rosette leaves of 5 week old plants and 150 ng of DNA used as input for each library. DNA shearing was carried out for 20 minutes at 37°C with 0.4U of DNA Shearase (Zymo research). The barcoded adapters used for library construction are listed in Rowan et al. Each set of 96 libraries was sequenced on one lane of an Illumina NextSeq500 instrument (300-cycle Mid Output run). Sequencing data was analysed to identify crossovers as previously reported, using the TIGER pipeline (Rowan et al; Yelina et al). These data can be compared to a set of wild type control libraries available at E-MTAB-4657. Rowan BA, Patel V, Weigel D, Schneeberger K. Rapid and Inexpensive Whole-Genome Genotyping-by-Sequencing for Crossover Localization and Fine-Scale Genetic Mapping. G3 (Bethesda). 2015;5: 385–98. Yelina NE, Lambing C, Hardcastle TJ, Zhao X, Santos B, Henderson IR. DNA methylation epigenetically silences crossover hot spots and controls chromosomal domains of meiotic recombination in Arabidopsis. Genes Dev. 2015;29: 2183–202.

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