title: GSM5333803: LacI WV1; Escherichia coli; OTHER
library_construction_protocol: Plasmids were prepped from each library using a Spin Miniprep Kit (Qiagen #27104). A nested amplicon PCR strategy was used to append barcodes and sequencing adapters. In a first round of PCR, amplicon primers were used to amplify 300 bp regions of the lacI gene and add standard Illumina MiSeq sequencing primer binding sites. The first six bases to be read in sequencing read 1 were degenerate to increase complexity and aid in base calling. In a second round of PCR, barcoding primers were used to append barcodes and flow cell adapter sequences. Amplicons were purified, quantified using Kapa Biosystems Library Quanitification kit (part number KK4824), and normalized. Libraries were sequenced on the Illumina MiSeq using MiSeq Reagent Kit v3, 600 cycle (part number MS-102-3003) according to the manufacturer's protocol.