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TransMessenger Transfection Reagent Handbook

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Introduction

TransMessenger Transfection Reagent has been designed to efficiently transfect eukaryotic cells with RNA, including siRNA (see page 14). This handbook contains guidelines and protocols for the transfection of single-stranded RNA (pages 913) and double-stranded siRNA (pages 1417).

The TransMessenger Principle
TransMessenger Transfection Reagent is based on a lipid formulation and is used in conjunction with a specific RNA-condensing reagent (Enhancer R) and an RNA-condensing buffer (Buffer EC-R). In the first step of TransMessengerRNA complex formation, the RNA is condensed by interaction with Enhancer R in a defined buffer system. In the second step, TransMessenger Transfection Reagent is added to the condensed RNA to produce TransMessengerRNA complexes. The TransMessengerRNA complexes are then mixed with serum-free medium and added directly to cells.

General Considerations for RNA Transfection

Transfection efficiencies are affected by a variety of parameters. The following factors should be considered carefully.

Cell culture
A healthy cell culture lays the foundation for successful transfection, particularly when transfecting cells with RNA. We strongly recommend subculturing cells for a minimum of 24 hours before transfection. This ensures normal cell metabolism and increases the likelihood of RNA uptake. Best results are obtained using the confluence levels indicated in the appropriate protocol sections. Contamination with bacteria (e.g., mycoplasma) and fungi should be avoided, since this can dra
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