title: Raw reads: 85-3 (ecotype_ID :22001)
design: To reduce starch, 3-week-old Arabidopsis thaliana accessions grown in soil were placed into darkness for 24-48 h before harvesting. 20-30 g of flash-frozen rosette tissue from pooled individuals were ground in liquid nitrogen with pestle and mortar. Nuclei isolation was performed as  described for the accession Ey15-2 (Rabanal et al., 2022; https://doi.org/10.1093/nar/gkac1115), and high-molecular-weight (HMW) DNA purified with Genomic-tips 100/G (Qiagen; #10243) following  manufacturer's instructions. 10 Âµg of HMW DNA were sheared with either Megaruptor 3 (Diagenode; #B06010003) or a needle (FINE-JECT 26Gx1" 0.45x25mm; #14-13651) to ca. 75 kb, and used as input for Continuous long reads (CLR) library preparation with the SMRTbell Express Template Preparation Kit 2.0 (Pacific Biosciences, California, USA; #101-693-800). These libraries were size-selected with the BluePippin system (SageScience) with a 30 kb cut-off in a 0.75% DF Marker U1 high-pass 30-40 kb vs3 gel cassette (Biozym; #BLF7510).
library_name: SMRTbell Express Template Preparation Kit 2.0