GSM8684651: DMSO rep 2 long-read; Arabidopsis thaliana; OTHER
Two leaves from each sample were flash-frozen in liquid nitrogen. Frozen tissue was ground into powder with a QIAGEN Tissuelyser II (30 beats/sec for 1min). High molecular weight DNA was extracted using a Promega Wizard HMW DNA Extraction Kit. DNA was sheared by passing through a 23G needle twice. Contaminating RNA and proteins were removed by treating with RNase A and Proteinase K (New England Biolabs). DNA was further cleaned and small fragments removed by performing a 0.7x bead cleanup using a modified SPRI bead mixture in which the buffer of Agencourt AMPure XP beads was exchanged with a custom buffer of 10 mM Tris-HCl, 1mM EDTA pH8, 1.6M NaCl, 11% w/v PEG 8000. Batch A was prepared using ONT Rapid Barcoding Kit V10 and sequenced on R9.4.1 flow cell. Batch B was prepared using ONT Native Barcoding Kit V14 and sequenced on R10.4.1 flow cell.