RNA-Seq of helianthus annuus roots 5 days post infestation with o. cumana
Whole roots were ground in liquid nitrogen, total RNA was isolated using SpectrumTM Plant Total RNA Kit (Sigma-Aldrich) according to the manufactures protocol. Libraries were prepared using the Genomics in house protocol for mRNA-seq. Briefly, the polyA fraction (mRNA) was purified from 500 ng of total RNA following by fragmentation and the generation of double-stranded cDNA. Then, end repair, a base addition, adapter ligation and PCR amplification steps were performed. Libraries were evaluated by Qubit (Thermo fisher scientific) and TapeStation (Agilent). Sequencing libraries were constructed with barcodes to allow multiplexing of the samples in 2 lanes. Approximately 16-20 million single-end 60-bp reads were sequenced per sample on Illumina HiSeq 2500 V4 instrument.
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