Illumina HiSeq 2000 paired end sequencing
Illumina 76bp paired-end RNAseq of root. Santander.
The RNASeq libraries were prepared from total RNA using the TruSeq RNA Sample Prep Kit v2 (Illumina Inc.,) with minor modifications. Briefly, 0.5_g of total RNA was used as the input material for after poly-A based mRNA enrichment with oligo-dT magnetic beads. The mRNA was fragmented (resulting RNA fragment size was 80-250nt, with the major peak at 130nt). After first and second strand cDNA synthesis the double stranded cDNA was end-repaired, adenylated and the 3'-T nucleotide of the adapter was used for the Illumina barcoded adapters ligation. The ligation product was enriched by 10 cycles of PCR.