GSM8187934: B1_AGY1613_Gen0_RepA; Saccharomyces cerevisiae; OTHER
Plasmids were recovered from each pool using Zymoprep Yeast Plasmid Miniprep II (Zymo Research D2004-A) with the following modifications: samples were incubated with 15 units zymolyase at 37°C for 1 hour, with inversion every 15 minutes; incubation in cell lysis buffer was extended to 10 minutes; after neutralization, samples were put on ice for 30 minutes, then centrifuged at 4°C. Plasmid barcodes were amplified using primers containing Illumina multiplex adaptors as described in (Piotrowski et al. 2015), with number of PCR cycles reduced to 20. Barcode amplicons were pooled and purified using AxyPrep Mag beads (1.8X volume beads per sample volume) according to the manufacturer's instructions (Axygen).