5.2. Using scAIR Positive Control RNA

The scAIR Positive Control RNA is designed for troubleshooting and measuring the sensitivity of the scAIR assay protocol. To run the positive control, aliquot 1 μl of scAIR positive control RNA in each well of a new 96-well plate with pre-aliquoted 5 μl of Hybridization Master mix. Freeze and store the plate at -80°C prior to use for the scAIR profiling experiment. For scAIR profiling, follow the standard protocol described below for experimental sorted T/B cells. To measure the sensitivity of the scAIR assay and mutation rate in amplified receptor sequences, you could consider adding spike-in TCR/BCR Synthetic RNAs (sold separately) to the positive control RNA. For sequences of spike-in TCR/BCR Synthetic RNAs, design of synthetic TCR/BCR control constructs, and spike-in control mixes, please refer to AIR Spike-in TCR/BCR Control Manual.

The scAIR Positive Control RNA included in the scAIR kits is RNA purified from CD3+ T cells or CD19+ B cells isolated from donor PBMC using CD3/CD19 antibody-magnetic beads.