Ampure xp clean up beads

mRNA was isolated with the RNA microkit (Qiagen) according to the manufacturer’s protocol. Isolated mRNA was used in the 5’ RACE-based SMARTer Mouse TCR α/β profiling kit (Takara Bio USA, Inc.) to perform sequencing of TCRs, following the manufacturer’s protocol using only the TCRβ-specific primers. Cleanup was performed with AMPURE XP clean-up beads (BD). PCR products were sequenced via Illumina MiSeq paired-end 2 × 300 nucleotide (nt) sequencing.

mRNA was isolated with the RNA microkit (Qiagen) according to the manufacturer’s protocol. Next, the 5′ RACE-based SMARTer Mouse TCR α/β profiling kit (Takara Bio, San Jose, CA, USA, Inc.) was used following the manufacturer’s protocol but only using the TCRβ-specific primers to perform TCR sequencing. The PCR products were cleaned up with AMPURE XP clean-up beads (BD) and sequenced via Illumina MiSeq paired-end 2 × 300 nucleotide sequencing.

mRNA was isolated with the RNA microkit (Qiagen) according to the manufacturer’s protocol. Isolated mRNA was used in the 5’ RACE-based SMARTer Human TCR a/b Profiling Kit v2 (Takara Bio USA, Inc.) to perform sequencing of TCRs, following the manufacturer’s protocol using only the TCRβ-specific primers. Cleanup was performed with AMPURE XP clean-up beads (BD). The resulting TCRβ libraries were sequenced on an Illumina MiSeq sequencer (paired-end 2x300nt). The reproducibility of the sequencing was analyzed by sequencing the libraries of donor 145 twice. A larger number of shorter reads was obtained for donor 204 and 292 on an Illumina NextSeq sequencer (paired-end 2x150nt) instead, although this did not lead to a dramatic increase in identified expansions (Figure 3A).