GSM5216200: PSO19_W12_CD8_S22_L003_R; Homo sapiens; RNA-Seq
Sequencing libraries were constructed using the SMART-Seq v4 Ultra Low Input RNA Kit (Takara Clontech cat. 634894). Bulk RNA was extracted from whole skin tissue using Qiagen RNeasy fibrous tissue mini kit (cat. 74704). Reverse transcription was performed on poly-adenylated RNA from cell lysate for up to 1,000 cells followed by second strand synthesis to make full length cDNA. cDNA was then later amplified by PCR using cycle numbers determined by cell numbers. Amplified cDNA was purified by AMPure XP beads (Beckman Coulter) and followed by Qubit quantification. Sequencing libraries were constructed using the Illumina Nextera DNA kit with 0.4 ng of amplified cDNA for sorted lymphocytes and 0.3 ng for cDNA synthesized from bulk skin RNA.