Whole genome sequencing of Saccharomyces cerevisiae:  dnCas3-CDA1_target 2
Genomic DNA was isolated from 5 replicates per strain using a standard zymolyase-based protocol. DNA concentrations were measured with a Qubit 2.0 and DNA quality was checked using a NanoDrop 8000 and by gel electrophoresis. Per sample, 1?g of genomic DNA was randomly fragmented by Covaris. Fragments with an average size of 200-400 bp were selected using the Agencourt AMPure XP-Medium kit. Fragments were end repaired and 3 adenylated. Adaptors were ligated to the 3 adenylated ends of each fragment and PCR amplified. PCR products were purified using the Agencourt AMPure XP-Medium kit. Purified PCR products were heat-denatured and circularized by a splint oligo sequence. The single-strand circular DNAs were quality-checked and used as the final library. Libraries were sequenced by BGISEQ-500 sequencing technology: through rolling-cycle replication of each single-strand circular DNA molecule a DNA nanoball (DNB) was formed, containing more than 300 copies of the initial circular DNA. The DNBs were loaded into a patterned nanoarray and reads (PE150) were obtained by combinatorial Probe-Anchor Synthesis (cPAS).