siRNA Transfection Reagent for Cell Lines From ThermoFisher

Stanford University
Biochemistry
Senior Research Scientist

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Company:

ThermoFisher Scientific

Product Name:

Lipofectamine RNAiMAX Transfection Reagent

Catalog Number:

13778075

Info: Supplier Page 

The WNT pathway is essential for embryonic development and adult tissue homeostasis. Deregulation of this pathway results in cancers of the stomach, intestine, breast, etc. I am interested in studying the molecular mechanism of oncogenic WNT signal transduction to identify potential targets for therapeutic intervention of colon cancer. In this experiment, the knockdown efficiency of an siRNA oligo pool targeting the LRP6 transcript in HEK 293T cells was tested.

Experimental Design and Results Summary

Application

Gene knockdown

Starting Material

HEK 293T cell line

Tips

Final concentration of siRNA was 25 nM. Knockdown duration was 72 hrs.

Results Summary

(a) 250,000 HEK 293T cells were seeded per well in a 6-well dish containing 2 mL of growth medium without antibiotics.(b) One hour post cell seeding, transfection with LRP6 siRNA (50 pmol siRNA + 5 µL Lipofectamine RNAiMAX transfection reagent) was done.(c) After 24 hours, medium was replaced with growth medium contains antibiotics. (d) 48 hours later, samples were harvested for Western blotting.(e) LRP6 immunoblot confirmed >95% knockdown efficiency.

DOI or PMID #

N/A

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Summary

The Good

siRNA transfection reagent that yields good knockdown efficiency.

The Bad

Higher amount of this transfection reagent might result in toxicity.

The Bottom Line

Works well for siRNA transfection in multiple cell lines.

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