|Transformationa||No. of transformants:||% of:|
|Ura+||Leu−b||LEU2/leu2 heterozygotesc||leu2Δ/leu2Δ homozygotesd|
|(−) Cas9, (+) vector||620||0||0||0|
|(−) Cas9, (+) gLEU2||700||0||0||0|
|(−) Cas9, (+) gLEU2, (+) repair fragment||650||0||0||0|
|(+) Cas9, (+) gLEU2||28||0||0||0|
|(+) Cas9, (+) gLEU2, (+) repair fragment||760||608||0||80|
↵a Transformations were performed with BWP17 (Cas9−) or HNY30 (Cas9+) with URA3-marked LEU2 gRNA plasmid, p499 (+gLEU2), or the vector that lacks gRNA (p494). (−), donor repair fragment absent; (+), donor repair fragment present. The results shown represent the average from 3 experiments.
↵b The number of Ura+ transformants that were leucine auxotrophs.
↵c The percentage of LEU2/leu2Δ heterozygotes was based on PCR analysis of LEU2 using genomic DNA isolated from 40 random Ura+/Leu+ transformants.
↵d The percentage of leu2Δ/leu2Δ homozygotes was based on PCR analysis of LEU2 amplified from genomic DNA isolated from 60 random Ura+/Leu− transformants.