title: GSM2967232: FL; Gossypium hirsutum; RNA-Seq
library_construction_protocol: Total RNA was extracted using an RNA Isolation Kit (Invitrogen™). The RNA was digested with RNase-Free DNase (Qiagen) and checked for integrity by capillary gel electrophoresis. Total RNA was separated into two parts. One was enriched using an oligo-(dT) primer to quantify the transcription levels of nuclear-encoded genes. The other portion of total RNA was subjected to ribosomal RNA depletion using the Ribo-Zero rRNA removal kit (Epicentre, USA) before cDNA library construction. The mRNA was enriched by oligo-(dT), then reverse-transcribed into cDNA with the SuperscriptTM first-strand synthesis system (Invitrogen, Carlsbad, CA, USA). The mRNA extracted by the Ribo-Zero rRNA removal kit was then fragmented into segments approximately 200 nucleotides in length and reverse-transcribed using random hexamer primers. The library was then constructed and sequenced using an Illumina HiSeqTM 2500. The raw sequence data were filtered and produced 8 Gb bases of clean reads