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. Author manuscript; available in PMC: 2013 Jul 26.
Published in final edited form as: Nat Protoc. 2012 Feb 2;7(2):374–393. doi: 10.1038/nprot.2011.446
Component Amount per tube Final
NEB ligation buffer, 10× 1 µl 1×
XhoI/EcoRI digested cTGM 100 ng 100 ng
XhoI/EcoRI digested 110-bp shRNA 4 ng 4 ng
NEB DNA ligase (2,000,000 U ml−1) 0.5 µl 1,000 U
DNase-free H2O Up to 10 µl