Skip to main content
. Author manuscript; available in PMC: 2009 Aug 4.
Published in final edited form as: Nat Protoc. 2009;4(3):341–355. doi: 10.1038/nprot.2008.250
Component Amount Final
5× ligase reaction buffer 4 µl 1×
ScaI → XmaI DNA fragment ends 30–120 fmol 30–120 fmol
BspEI → ScaI DNA fragment ends 30–120 fmol 30–120 fmol
T4 DNA ligase (1 U µl−1) 1 µl 1 U
Nuclease-free water Up to 20 µl