RNA-seq of Glycine max: root at 15 days old
Soybean roots were flash frozen in liquid nitrogen. RNA was extracted from four biological replicates using CTAB method (Chang et al., 1993) and purified in acidic phenol/chloroform. RNA was treated with DNase I (New England Biolabs), precipitated with isopropanol and re-suspended in RNase-free water. Then, RNA concentration and quality was measured using Nanodrop and Bioanalyzer. RNAseq libraries were prepared with Illumina's 'TruSeq Stranded mRNAseq Sample Prep kit' (Illumina). The libraries were quantitated by qPCR and sequenced on one lane for 101 cycles from each end of the fragments on a HiSeq2500 using a HiSeq SBS sequencing kit version 4. Fastq files were generated and demultiplexed with with the bcl2fastq v2.17.1.14 Conversion Software (Illumina).