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PLZF knockdown using siRNA

SA Shuchi Agrawal Singh

Last updated date: May 22, 2023 Views: 535 Forks: 0

original An abbreviated version of this protocol was published in eLife in Jan, 2019
PLZF targets developmental enhancers for activation during osteogenic differentiation of human mesenchymal stem cells
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Transient transfection in human mesenchymal stem cells.

 

 Reagents:
1) 6uM siRNA per 1 million cells
2) Optimem
3) Lipofectamine 2000
Protocol:
tube 1) 500ul Optimem + 6ul siRNA (100uM stock)
tube 2) 500ul Optimem + 10ul Lipo2000
mix tube 1 and 2
incubate 20min RT
3) Add transfection complex to 1million cells pellet. Mix well and
4) plate in 10cm dish with hMSC media.

Next day change media and harvest cells at appropriate time point.

 

Hope this helps


Good Luck
Kind regards
Shuchi

Copyright: © 2023 The Author(s); This is an open-access article under the CC BY-NC license (https://creativecommons.org/licenses/by-nc/4.0/).
How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Agrawal Singh, S(2023). PLZF knockdown using siRNA. Bio-protocol Preprint. bio-protocol.org/prep2310.
  2. Agrawal Singh, S., Lerdrup, M., Gomes, A. R., van de Werken, H. J., Vilstrup Johansen, J., Andersson, R., Sandelin, A., Helin, K. and Hansen, K.(2019). PLZF targets developmental enhancers for activation during osteogenic differentiation of human mesenchymal stem cells. eLife. DOI: 10.7554/eLife.40364

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