GSM6926793: 620_RS_11; Mus musculus; OTHER
Sorted cells were stored at -80C. RNA was extracted from frozen cells using the RNeasy Plus Micro Kit. TCRβ amplicon libraries were generated with 5'Rapid Amplification of cDNA Ends (5'RACE) technology. RNA was transcribed to cDNA with a TCRβ constant region primer and a modified SMARTerIIA primer (Integrated DNA Technologies), with 9nt long unique molecular identifiers (UMI) added to individual TCRβ cDNA sequences. TCRβ locus was amplified by nested PCR with an additional TCRβ constant region primers, and a universal primer to SMARTerIIA, with the final PCR adding sequencing adaptors and barcodes to the TCRβ libraries. Products after each PCR were purified using AMPure XP AgenCourt Beads (Beckman Coulter).