GSM8743048: Figure 2,  Donor 002,  CD22_targeted_long_linker_CAR_with_CD28-CD3z_signaling_domain (28l), Lane 3,  biol rep 1; Homo sapiens; RNA-Seq
RNA was harvested from 1 million CAR positive Primary T cells using Quick-RNA™ Microprep Kit” from zymo research. 10ng of total RNA was used for creation of sequencing libraries. Total RNA integrity was determined using Agilent Bioanalyzer or 4200 Tapestation. Library preparation was performed with 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.