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. Author manuscript; available in PMC: 2017 Oct 1.
Published in final edited form as: Nat Protoc. 2017 Mar 23;12(4):828–863. doi: 10.1038/nprot.2017.016
Steps Cloning process Number of reactions
3–7 PCR amplification of pooled oligo library 12
8–10 Restriction digest of plasmid backbone 16
11–12 Gibson assembly 10 with sgRNA insert, 5 control
13–17 Isopropanol precipitation 10 with sgRNA insert, 5 control