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Large insert-size library of O. sativa was created according to the manufacturer’s protocols (Oxford Nanopore, UK). Briefly, 5 µg genomic DNA was sheared into ~20-30 kb fragments using g-TUBE (#520079, Covaris) centrifugation (twice at 1,400 g for 2 min) and size-selected (>8-10 kb) by Blue Pippin (Sage Science, MA) using a marker started at 5-12 min (0.75% DF Marker S1 High-Pass 6-10kb vs3) to ensure the removal of small DNA fragments. Genomic DNA libraries were prepared using a Ligation sequencing 1D kit (SQK-LSK109, Oxford Nanopore, UK). End-repair and dA-tailing of DNA fragments were performed using an Ultra II End Prep module (#E7546, NEB) according protocol recommendations. Each dA-tailed sample was tethered to 1D adapter using a Quick Ligation Module (#E6056, NEB). The prepared DNA library was loaded into a FLO-PRO002 flow cell and sequenced on PromethION sequencers (Oxford Nanopore, UK). The raw data collected in this experiment was obtained as fast5 files after conversion of electrical signals into base calls via guppy 2.0.8 (Oxford Nanopore, UK). |
WGS |
GENOMIC |
unspecified |
SINGLE
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