title: GSM5420676: TCR_1_4_engineered_T_cells_scRNA-seq; Homo sapiens; RNA-Seq
library_construction_protocol: Chromium next GEM Single Cell VDJ V1.1, Rev D; In order to amplify transgenic TCR sequences, the cDNA was amplified in two steps in parallel to the VDJ enrichment by using the 10x forward outer and inner primers as well as TCR- and/or editing method-specific reverse primers (see Supplementary Table 2). In target enrichment 1 and 2 (protocol step 4.1 and 4.3) the elongation time was increased to 90 seconds in adaptation to the longer transduction products. In addition, the second amplification of the KI constructs was performed for 15 cycles instead of 10 in order to compensate for lower integration frequencies. The double-sided size selection (Step 4.4 of the 10x protocol) was adjusted to 0.4X and 0.8X for the transduction constructs and was kept at 0.5X and 0.8X for the KI constructs. Chromium™ Next GEM Single Cell 5' Library & Gel Bead Kit