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TransMessenger Transfection Reagent Handbook

followed throughout the procedure.

General handling
Proper microbiological, aseptic technique should always be used when working with RNA. Hands and dust particles may carry bacteria and molds, and are the most common sources of RNase contamination. Always wear latex or vinyl gloves while handling reagents and RNA samples to prevent RNase contamination from the surface of the skin. Change gloves frequently and keep tubes closed. Ensure that all glassware, plasticware, and solutions (including water and culture medium) are RNase-free.

Disposable plasticware
The use of sterile, disposable, plastic tubes throughout the procedure is recommended. These tubes are generally RNase-free and do not require pretreatment to inactivate RNases.

DNA contamination
No currently available purification method can guarantee that RNA is completely free of DNA, even when no DNA is visible on an agarose gel. For RNA transfection, digestion of the purified RNA with RNase-free DNase is recommended. The RNA can then be repurified using the RNeasy Kit.

Optimizing RNA Transfection

To achieve optimal transfection efficiency for every cell typeRNA combination, we recommend optimizing a number of parameters:

The amounts of TransMessenger Transfection Reagent, RNA, and TransMessengerRNA complexes
The cell number/confluency
The length of exposure of cells to TransMessengerRNA complexes

Once the parameters yielding maximum transfection efficiency have been determined, they should be kept constant in every experiment usi
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