GSM8646266: Atsdr4l-5 dig2, seed coat, rep 3; Arabidopsis thaliana; RNA-Seq
Seeds were gently pressed between two microscope glass slides to break the seed coats. Endosperm (with seed coat) and intact embryos were manually separated using syringe needles and fine forceps and preserved in RNAlater buffer (AM7020, Thermo Fisher Scientific, Waltham, MA, USA) in a microcentrifuge tube until all samples have been collected. Total RNA was isolated using Spectrum™ Plant Total RNA Kit (STRN50, MilliporeSigma, Burlington, MA, USA), followed by the removal of genomic DNA by DNase I treatment (AM1907, TURBO DNA-free™ Kit, Thermo Fisher Scientific, Waltham, MA, USA). NEBNext® Ultra II Directional RNA Library Prep Kit for Illumina® (E7760S, NEB, Ipswich, MA, USA) with NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) and Unique Dual Index UMI Adaptors RNA Set 1 (E7416S, NEB, Ipswich, MA, USA) were used for library preparation following the manufacturer's instructions.