Diluted PIBv1 MPRA plasmid library with GFP ORF amplified alongside cDNA of Jurkat replicate 1, sequencing lane 4
Amplicon library of MPRA plasmid library (with GFP ORF) for contrast with tag sequencing of GFP cDNA (i.e. to determine if the oligo results in active reporter). Following Tewhey et al. 2016, amplicons were generated by PCR with custom primers, then the product was diluted to equal the concentration of the amplified cDNA library from the same cell type with the lowest concentration, followed by 6 further cycles of PCR to add Illumina adapters. The 6 cycles of PCR for Illumina adapter addition were performed simultaneously for diluted plasmid library and the corresponding cDNA library from each biological replicate, so each mpra:gfp library is paired with a corresponding cDNA library.
SUB16055622
PIBv1_mpragfp_Jurkat_rep1_L004