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. Author manuscript; available in PMC: 2019 May 13.
Published in final edited form as: Nat Protoc. 2018 Nov;13(11):2615–2642. doi: 10.1038/s41596-018-0055-0
Component Volume (μl) Final concentration
T4 DNA ligase buffer (10×) 1 1×
BsaI-digested vector backbone 20 ng variable 2 ng/μl
Diluted oligoduplex from (iii) 2 2 nM
T4 PNK (10 U/μl) 0.5 0.5 U/μl
T4 DNA ligase (400 U/μl) 0.5 20 U/μl
Nuclease-free H2O to 10
Total 10