Component	Amount
Double gRNA plasmid (7f)	1 μg#
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

^*

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

^#

When transforming multiple plasmids, we recommend using 2–5 μg of DNA per plasmid to increase the number of transformants.