Component	Amount
pMEL plasmid backbone (3.a.i.1–2 & 3b)	100 ng
gRNA insert fragment (2.a.3)	500 ng
Double stranded repair DNA (4)	*
50% PEG	240 μL
Lithium acetate (1M)	36 μL
SSDNA	25 μL
Distilled water (sterile)	X μL
Total volume	351 μL

^*

When aiming for gene deletions or the introduction of SNPs, we recommend using 1 μg of repair DNA. When aiming for (multiple) gene integration, we recommend using at least 200 ng/kb for each fragment.