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. Author manuscript; available in PMC: 2010 Apr 22.
Published in final edited form as: Nat Protoc. 2009 Sep 17;4(10):1471–1501. doi: 10.1038/nprot.2009.98
Component Amount Final Concentration
SapI- and Pfu-treated pKJ1712 backbone 2 μl ∼1 ng μl-1
Annealed binding site oligos (or water for control) 8 μl 19 nM
2× Quick Ligase Buffer 10 μl 1×
T4 DNA Ligase (400 U μl-1) 1 μl 19 U μl-1
Total 21 μl