GSM4804935: HT_3W_3; Solanum tuberosum; RNA-Seq
Upper 5 leaves were harvested for RNA isolation. Total RNA were extracted from 50 mg of frozen ground tissues using RNeasy Plant Mini kit (QIAGEN). Five micrograms of each RNA sample were used to generate a strand-specific library containing inserts of approximately 150–200 bp in size. PolyA RNA is captured by oligodT beads (Thermo Scientific, USA) and fragmented. The first-strand cDNA as synthesized by Superscript III reverse transcriptase (Invitrogen,USA) using dNTP, whereas the second-strand cDNA was generated using a dUTP mix. After end-repair, dA-tailing and adapter ligation, the dUTP-containing second strand is digested by uracil DNA glycosylase (Enzymatics, USA). Indexed cDNA libraries were prepared by PCR amplification.