Component	Amount (μL)
FastDigest Green Buffer (10x)	1
Plasmid (6.b.v)	250 ng
FastDigest Enzyme(s)**	0.5*
MilliQ	X
Total volume	10

^*

When multiple restriction enzymes are used, add 0.5 μL of each enzyme.

^**

When each gRNA coding sequence contains a restriction site, two separate reactions should be performed where only one of the restriction enzymes corresponding to the gRNA restriction sites is added.