title: Gossypium barbadense Transcriptome Sequences
design: The seed of Gossypium barbadense L.(AD)2, genetic standard line 3-79, Gossypium tomentosum (AD)3, Gossypium mustelinum (AD)4, Gossypium armourianum (D2-1), and Gossypium longicalyx (F) were planted at Texas A&M University. Young leaf tissues were sampled from each plant and used to isolate total RNA using the Qiagen RNeasy Mini Kit per manufacturer instructions. RNA isolates were quantified using NanoDrop spectrophometry and checked for quality by gel electrophoresis. Illumina TruSeq RNA libraries were prepared using the manufacturer protocol. Libraries were normalized (Matvienko et al. 2013) by denaturation and rehybridization in NaCl and TMAC buffers and were then treated with Duplex Specific Nuclease to digest remaining DNA contamination (Zhulidov et al. 2004). The treated library was then re-amplified using the Illumina library primers via PCR per manufacturers instructions. The libraries were then sequenced using the Illumina Genome Analyzer II for 85 cycles.