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. Author manuscript; available in PMC: 2018 Nov 1.
Published in final edited form as: Nat Protoc. 2018 Apr 5;13(5):875–898. doi: 10.1038/nprot.2018.007
Component Amount Final concentration
pSMART-sgRNA vector 10 µg 100 ng/µl
10× Buffer 2.1 10 µl 1×
BbsI (10 U/µl) 5 µl 0.5 U/µl
Nuclease-free H2O Make up to 100 µl