Component	Amount (μL)
DreamTaq Master Mix (2x)	50
pROS template*	1–100 ng
gRNA (Ptarget ROS1)(10 μM)	2
gRNA (Ptarget ROS2) (10 μM)**	2
MilliQ	X
Total volume	100

^*

This can be any pROS plasmid

^**

When aiming to construct a pROS plasmid targeting a single distinct locus, only gRNA primer 1 is used.