I was wondering if anyone has already experience with constructing sgRNA libraries. We are interested in doing a knock-out experiment (loss-of-function) with 5 different conditions. I have found this protocol from Joung et al.
In the protocol they have 10 different Fwd primer with no unique barcodes, only the eight Rev primers contains unique barcodes for different samples in KO.
Do I understand it correctly, that i can pool together only eight samples in one sequencing run with this protocol?
Are there more complex library preparation kits out there which can deal with more samples?