Characterization of distinct T cell receptor repertoires in tumor and adjacent non-tumor tissues from lung cancer patients

We collected tumor tissues and matched non-tumor tissues from 15 lung cancer patients and RNA was extracted from these specimens. Then, RNA was applied to prepare libraries of TCR. A series of three nested PCR reactions as described previously (PMID: 24952902) were performed to obtain a TCR repertoire; the ARM-PCR procedure (PMID: 20080641) was slightly modified in this assay. After complete three nested PCR reactions, DNA with lengths of 200-500 bp was separated and purified with a QIAquick gel extraction kit (Qiagen, cat. no. 28706). Amplicons were then sequenced.