GSM4565821: Multiple libraries 1; synthetic construct; OTHER
Zymoprep was used to extract the plasmid containing the gene for the peptide-MHC. Peptide library was encoded in an appropriate oligo using degenerate oligos (e.g. NNK) where appropriate and as described in the manuscript. The naive library and round 1, 2 and 3 were incubated with magnetic-streptavidin beads coated with TCR of interest at 400 nM. After incubation for 3 hours at 4C, the fraction of TCR bound beads was isolated using LS magnetic columns (Miltenyi). For round 4 of the Pmhc selection a TCR-streptavidin tetramer was used iin place of the streptavidin-beads.
GEO Accession
GSM4565821