GSM8766409: PBS-inoculated TIGK cells, biological replicate 2; Homo sapiens; RNA-Seq
RNA was extracted from whole cells using a Trizol-based protocol, genomic DNA was digested using TURBO DNaseI according to manufacters protocol and samples were cleaned using Qiagen RNeasy kit. Total RNA was used for the construction of sequencing libraries. Libraries were prepared using the Zymo-Seq RiboFree Total RNA Library Prep Kit (Cat # R3000) according to the manufacturer's instruction manual v1.3.0 RNA was reverse-transcribed into cDNA, which was followed by ribosomal RNA depletion. After that partial P7 adapter sequence was ligated at 3' end of cDNAs, followed by second strand synthesis and partial P5 adapter ligation to 5' end of the double stranded DNAs.  Lastly, libraries were amplified to incorporate full length adapters under the following conditions: initial denaturation at 95°C for 10 min; 10 - 16 cycles of denaturation at 95°C for 30 sec, annealing at 60°C for 30 sec, and extension at 72°C for 60 sec; and final extension at 72°C for 7 min