Simultaneous knockdown of the expression of two genes using multiple shRNAs and subsequent knock-in of their expression

. Author manuscript; available in PMC: 2009 Sep 28.

Published in final edited form as: Nat Protoc. 2009 Aug 27;4(9):1338–1348. doi: 10.1038/nprot.2009.145

Component	Volume per reaction (μl)	Final concentration or amount
10× reaction buffer	5 μl	1 × reaction buffer
plasmid from step 18 (variable concentration)	variable	1–5 μg
each restriction enzyme (10–20 μ 10³ units/ml)	variable	2–5 units/ μg of DNA
ddH₂O	variable	To a final volume of 50 μl