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TransMessenger Transfection Reagent Handbook

Influence of the RNA Factors such as the size of the RNA and special features contained within it (e.g., IRES, cap, or poly-A tail) can influence its stability and/or the expression rate. Some RNA features may not be compatible with every cell type. Sub-optimal cell
density If cell density at the time of TransMessenger RNA complex addition is not at an optimal
level, cells may not be in the optimal growth phase. This can lead to insufficient uptake of
complexes into the cells or inefficient processing of the RNA of interest. For adherent cells, the optimal confluency for transfection is 8090% (for single-stranded RNA) or 5080% (for siRNA). Be sure to seed cells a minimum of 24 h prior to transfection. Cell typeRNA
combination RNA species with features such as a cap, poly-A tail, or IRES will not necessarily be processed in every cell type. For example, an RNA with an IRES may not be translated in COS-7 and HeLa cells. If no expression is observed change the cell type or, if possible, the RNA. Poor RNA quality RNA used for transfection should be of high quality. Impurities present in RNA preparations can potentially lower transfection efficiency. Degradation of RNA leads to decreased expression levels. RNA should be checked for degradation before transfection. We recommend purifying RNA using RNeasy Kits, Oligotex mRNA Kits, or QIAGEN DNA/RNA Kits. Rep
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