title: Cryptic-Seq: Bxb1 attP-GT rep 3
design: HEK293attB gDNA was incubated with Tn5 transposase and transfected with donor plasmid that includes an attP-GT integrase attachment, and Bxb1 at 37C for 4h in a recombination buffer. PCR was used to amplify the regions of integration using Q5 polymerase (NEB) and primers for P5 containing distinct barcodes for each sample (LM_P5_F1-F3) and a single N7 primer (GN037_CrypticSeq_N7_i7_N701). Following PCR, a 1.5x AMPure bead clean-up was performed, and products were run on a Tapestation 4200 (Agilent Technologies) using D1000 tape to confirm amplification. Library concentration was determined using the Next Library Quant Kit (NEB) for Illumina. Libraries were normalized to 2nM, loaded at a concentration of 750pM and sequenced via Illumina Next Seq according to the manufacturers instructions.