Amplicon sequencing of plasmid library: N80 liquid replicate 2
Sequencing library was prepared by PCR with NEBNext Ultra II Q5 Master Mix (NEB M0544) in two consecutive reactions. The barcode region of the plasmid library was amplified and flanked with Illumina TruSeq read 1 and read 2 sequences by PCR. The library was then indexed using custom ordered TruSeq indexing primers in a second PCR and purified by freeze-and-squeeze gel extraction followed by SPRI bead purification.