Identifying crossover locations from pollen of Arabidopsis thaliana Col x Ler F1 plants (WT and recq4) using Nanopore-seq and COmapper

ABSTRACT: We sequenced pollen DNA from Col x Ler F1 hybrid plants using Nanopore long-read sequencing and identified crossover sites with COmapper. To purify pollen grains from F1 hybrid plants and extract their genomic DNA, whole inflorescences of hybrid plants were collected and transferred to ice-cold 10% (w/v) sucrose in a 50-ml tubes. Flowers were homogenized with a blender (Tefal, BL3051KR), filtered through 80-μm nylon mesh, and centrifuged at 350g for 10 min at 4 °C. The supernatant was discarded and the pellet was resuspended in 10% (w/v) sucrose, filtered through a 40-μm cell strainer, and centrifuged at 100g for 10 min at 4°C. The purified pellet was frozen in liquid nitrogen and ground with a mortar and pestle. The pellet powder was resuspended in CTAB buffer (1% [w/v] CTAB, 50 mM Tris-H

INSTRUMENT(S): PromethION

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Kyuha Choi 

PROVIDER: E-MTAB-14369 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress