Ion torrent pgm 316 or 318 chip

DNA barcode amplification was performed using Taq PCR Master Mix Kit (Qiagen, 201443) and 3 μm of forward and reverse primers (LGC Biosearch Technologies)^{22 (link)}. PCR amplification was conducted on sorted multimer-binding T cells (in < 20 μL of buffer) and on the stored aliquot of the MHC multimer reagent pool (diluted 50,000 × in the final PCR) under the following conditions: 95 °C 10 min; 36 cycles: 95 °C 30 s, 60 °C 45 s, 72 °C 30 s, and 72 °C 4 min. The multimer reagent pool was used as the baseline to determine the number of DNA barcode reads within a non-processed MHC multimer reagent library. PCR products were purified with a QIAquick PCR Purification kit (Qiagen, 28104) and the amplified DNA barcodes were sequenced at Sequetech (USA) using an Ion Torrent PGM 316 or 318 chip (Life Technologies).

DNA barcode amplification was performed using Taq PCR Master Mix Kit (QIAGEN, 201443) and 3 μM of forward and reverse primers (LGC Biosearch Technologies). PCR amplification was conducted on sorted multimer-binding T cells (in <19 μL of buffer) and on a triplicate of the stored aliquot of the MHC multimer reagent pool (diluted 10.000× in the final PCR) under the following conditions: 95°C for 10 minutes; 36 cycles: 95°C for 30 seconds, 60°C for 45 seconds, 72°C 30 for seconds, and 72°C for 4 minutes. The multimer reagent pool was used as the baseline to determine the number of DNA barcode reads within a nonprocessed MHC multimer reagent library. PCR products were purified with a QIAquick PCR Purification Kit (QIAGEN)m and the amplified DNA barcodes were sequenced at PrimBio using an Ion Torrent PGM 316 or 318 chip (Life Technologies).