Bio-protocol.org
11. Perform Gibson assembly by mixing 50 ng digested pCI-neo, 50 ng left arm, 50 ng right arm, 85 ng GFP-mCherry (the molarity ratio of pCI-neo and other three fragments are 1:5:5:5) in 10 ml Gibson Assembly® Master Mix (NEB, E5510) and double-distilled H. 2 O to 20 ml. Incubate at 50°C for 30 min. ................
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