GSM8809493: CTCL1024_CD4P_CD7N_OW; Homo sapiens; RNA-Seq
RNA was extracted from skin, lymph node, and purified PBMC samples processed as described above using the RNeasy Plus Mini kit (Qiagen). Libraries were prepared by the WU GTAC@MGI using 10ng of total RNA with a Bioanalyzer RIN score greater than 8.0. ds-cDNA was prepared using the SMARTer Ultra Low RNA kit for Illumina Sequencing (Takara-Clontech) per manufacturer's protocol. cDNA was fragmented using a Covaris E220 sonicator using peak incident power 18, duty factor 20%, cycles per burst 50 for 120 seconds. cDNA was blunt ended, had an A base added to the 3' ends, and then had Illumina sequencing adapters ligated to the ends. Ligated fragments were then amplified for 12-15 cycles using primers incorporating unique dual index tags. Fragments were sequenced on an Illumina NovaSeq-6000 using paired end reads extending 150 bases. A subset of samples had previously been subjected to RNAseq using the following methods: Takara Bio SMARTer cDNA synthesis followed by library preparation according to manufacturer's protocol (Takara), indexed, pooled, and sequenced (1x150 bp SE) on an Illumina HiSeq 3000 instrument according to the manufacturer's instructions.