RNA-Seq of Lotus japonicus kai2akai2b
Root tissue from kai2akai2b was harvested and snap-frozen in liquid nitrogen. RNA was extracted using Spectrum Plant Total RNA Kit (Sigma-Aldrich, USA). DNA contamination was removed with an on-column DNAse I treatment. Residual DNA was further removed by LiCl precipitation. RNA purity was tested by PCR to verify absence of genomic DNA. Subsequently, RNA quality was tested on an Agilent 2100 Bioanalyzer (Agilent Technologies, USA), and samples with RIN>6.7 were processed further. Library construction was carried out with TruSeq Stranded mRNA LT (RS-122-2101, Illumina, USA) after selection with AMPure XP beads (NEB). Sequencing was performed on an Illumina HiSeq 2500 (Illumina, USA) to obtain 100bp paired-end reads.