you mentioned that an ideal sgRNA library should have >70% perfectly matching guides, <0.5% undetected guides, and a skew ratio of less than 10.
How did you reach these numbers?
I made 4 costume libraries . Undetected guides in all are less than 5 (0.2). The skew ratio is less than 10 (4.3-5.6). But the percentage of matched guides is 68%, 68%, 72% and 82 % for library 1 to 4 respectively.
Do you think I should remake the library 1 and 2 ?
I have no idea what to do at this step. I highly appreciate your opinion .