Overexpression of Mj-Ise6,Mj-MSP400,Mj-Nulg1A,Mj-Nulg1B and GFP in S. lycopersicum hairy roots (mRNA-sequencing)
Two weeks after growing tomato hairy in liquid mediumMj-Ise6,Mj-MSP400,Mj-Nulg1A,Mj-Nulg1B and GFP expression were induced with Estradiol for 24 h. For mRNA sequencing, RNA was extracted from 100 mg grounded tissue using the RNeasy Plant Mini Kit, Qiagen. Four biological replicates, each independently transformed, were used per treatment. RNA quality control was performed. RNA 6000 nanochip, Agilent technologies libraries were prepared using QuantSeq 3' mRNA library prep FWD kit , Lexogen according to the manufacturers instructions. Library quality was verified using a High sensitivity DNA chip, Agilent technologies. Library quantification was assessed using qPCR assay according to Illumina protocol equimolar pooling of libraries based on qPCR and libraries were sequenced on an Illumina NextSeq 500 platform, SR76, with high output. The sequencing depth was 8M.