title: GSM3332222: EB4_Kcnh2_WT rep2; Rattus norvegicus; RNA-Seq
library_construction_protocol: When rat embryonic stem cells (WT, Kcnh2-/-) form embryoid body and differentiate to day 4, total RNA was extracted through RNAiso Plus (Takara).  In order to obtain high-quality sequencing data, ribosomal RNA was removed using a biotin-labeled specific probe (Ribo-ZeroTMrRNA Removal Kit). After purification, the RNA was fragmented under a certain temperature and ionic environment. Subsequently, using the random primers and reverse transcriptase in the TruSeq® Stranded kit synthesizes cDNA strands. And then double-stranded cDNA was synthesized using DNA polymeraseⅠ and RNase H. During this progress, the RNA template was removed and dTTP was replaced by dUTP. The double-stranded cDNA was treated with the addition of “A” bases and adaptors. After amplification and purification, the cDNA library was obtained. Finally, the constructed cDNA library was sequenced using Illumina Hiseq platform with the 100-bp pair-end sequencing strategy. RNA libraries were prepared for sequencing using standard Illumina protocols