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TransMessenger Transfection Reagent Handbook

te to ensure uniform distribution of the transfection complexes.
Note:
Use of media without serum or antibiotics avoids the potential introduction of RNases. It is possible that transfection in the presence of serum may give improved results. Double-stranded siRNA is more resistant to degradation by RNAses than singlestranded RNA and therefore, if RNase contamination is not a concern, or if your serum is validated to have no detectable RNase activity, medium containing serum can be used. 9. Incubate cells with the transfection complexes for 3 h under their normal growth conditions.
Note:
If no cytotoxic effects are observed, the incubation time can be increased up to 4 h. 10. Remove complexes from the cells, wash cells once with PBS, and then add 500 l fresh medium containing serum and antibiotics to the cells. 11. Incubate cells under their normal growth conditions and monitor gene silencing after an appropriate incubation time. Change medium as required.
Note:

The optimal time point for gene silencing analysis is dependent on cell type, the gene targeted, and method of analysis. A time course experiment should be performed to determine the appropriate incubation time.

Troubleshooting Guide

The following troubleshooting guide is helpful if lower transfection efficiencies, hig
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DNase I (Deoxyribonuclease I) digests single- and double-stranded DNA to oligodeoxyribonucleotides. Ribonuclease activity has been reduced to non-detectable levels....
... coli containing a clone of the human Topoisomerase ... II alters the topological state of nucleic acids ... transient break which generates a separate DNA helix ... DNA passage mechanism, the enzyme can relax negatively ...
RNase-Free DNase 50 l...
supplied with 10x reaction buffer...
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