Transformation outcomes

ConstructTransformation no.aRecipient strainbIntroduced geneNo. of transformants recovered
sgRNA1sgRNA2Split-marker repair templateTotalHisLeu
UME6 deletion1SN1520000
3SN152UME6 (1 µg)0ume6Δ::r1HIS1r1333
BRG1 deletion and HIS1 excision in ume6Δ::r1HIS1r14MH1010000
6MH101BRG1 (1 µg)0brg1Δ::r2LEU2r21,1200
7MH101BRG1 (1 µg)HIS1 (1 µg)brg1Δ::r2LEU2r256422
8MH101BRG1 (1 µg)HIS1 (3 µg)brg1Δ::r2LEU2r227642
9MH101BRG1 (1 µg)HIS1 (9 µg)brg1Δ::r2LEU2r215630
BCR1 deletion and LEU2 excision in ume6Δ::r1 brg1Δ::r2LEU2r210MH1100000
12MH110BCR1 (1 µg)0bcr1Δ::r1HIS1r11340
13MH110BCR1 (1 µg)LEU2 (1 µg)bcr1Δ::r1HIS1r14722
  • a All transformations included a CAS9 gene, following the method of Min et al. (10). Approximate amounts of each PCR product in a typical transformation were the following (unless otherwise stated within the table): CAS9, 1 µg; sgRNA1, 1 µg; sgRNA2, 1 µg; split-marker cassette A, 1.5 µg; split marker cassette B, 1.5 µg.

  • b All strains are of genotype his1Δ/his1Δ leu2Δ/leu2Δ arg4Δ/arg4Δ. MH101 has the additional genotype ume6Δ::r1HIS1r1/ume6Δ::r1HIS1r1. MH110 has the additional genotype ume6Δ::r1/ume6Δ::r1 brg1Δ::r2LEU2r2/brg1Δ::r2LEU2r2, in which the ume6Δ::r1 allele is marked only with one copy of the flanking repeat from the r1HIS1r1 marker cassette.