Simple, robust strategies for generating DNA-directed RNA interference constructs.

  title={Simple, robust strategies for generating DNA-directed RNA interference constructs.},
  author={Robert R. Rice and A. Muirhead and Bruce Harrison and A. Kassianos and P. Sedlak and N. Maugeri and P. J. Goss and J. Davey and David E James and Michael W Graham},
  journal={Methods in enzymology},
We describe two complementary strategies for preparing DNA-directed RNA interference (ddRNAi) constructs designed to express hpRNA. The first, oligonucleotide assembly (OA), uses a very simple annealing protocol to combine up to 20 short nucleotides. These are then cloned into appropriately designed restriction sites in expression vectors. OA can be used to prepare simple hairpin (hp)-expressing constructs, but we prefer to use the approach to generate longer constructs. The second strategy… Expand
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