Plant sample from Hordeum vulgare
For each large-insert size mate-pair library, 20–60 μg of genomic DNA was sheared to the desired insert size using nebulization for 2 kilobases (kb) or HydroShear for 6 kb. DNA fragments were biotin-labeled, size selected, and circularized. Circular DNA molecules were sheared with Adaptive Focused Acoustic (Covaris) to an average size of 450 bp. Biotinylated fragments were purified on magnetic beads (Invitrogen) and used to construct libraries.