Black wheat
1ug of isolated total RNA samples were used forsequencing library preparation using Kapa mRNAhyperprep Kit (KR1352  v5.17) according to themanufacturers instructions.Briefely, mRNA was captured using magnetic oligo-dT beads from 300ng of intact total RNA followed byheat and magnesium induced fragmentation.First strand cDNA synthesis of the fragmentedmRNA was performed using random priming whichis followed by combined second strand synthesisand A-tailing, which converts the cDNA:RNA hybridto double-stranded cDNA (dscDNA), incorporatesdUTP into the second cDNA strand, and adds dAMPto the 3' ends of the resulting dscDNA.Then dsDNA adapters with 3' dTMP overhangs wereligated to library insert fragments along withsubsequent limited cycle high-fidelity libraryamplification. During amplification the strandmarked with dUTP is not amplified, allowing strand-specific sequencing.Quality of RNA-Seq libraries were checked usinghigh sensitivity D1000 ScreenTape in Agilent 2200TapeStation system and final library quantificationwas done using Qubit (Thermo Scientific). Pairedend 2 x 150 bp sequencing of these libraries wereperformed in Novaseq 6000 (Illumina).