Cells to ct kit manual

The TaqMan® Cells-to-CT™ Control Kit was designed for use with TaqMan® Cells-to-CT™ products. This kit contains an endogenous control (ACTB) to normalize for sample loading variability and an artifical XenoRNA™ control and corresponding TaqMan® Gene Expression assay to monitor the effects of PCR inhibition. Procedure overview The TaqMan MicroRNA Cells-to-CT Kit procedure is shown in Figure 1 and described below. • First, 10– cultured cells are washed with phosphate-buffered saline (PBS), mixed with Lysis So lution, and incubated at room tem-perature for 8 min. Cells are lysed during this incubation and RNA. The protocols for manually processing cells with Cells-to-CT Kits recommend mixing samples after the addition of both the the Lysis and Stop Solutions. To facilitate automation of the procedure, we investigated whether these sample mixing steps are required for reproducible, high quality real-time PCR data using the TaqMan Gene Expression Cells-to-CT Kit.

The Cells-to-CT™ Kits for TaqMan® and SYBR® Green-based real-time PCR take you from cells in culture to real-time RT-PCR analysis without RNA purification. The Cells-to-CT system uses a proprietary method for lysing cultured cells while simultaneously removing genomic DNA and preserving RNA integrity, saving significant time and effort over traditional methods requiring RNA purification. Cell type compatibility Table 1 lists the cell types that have been shown to be compatible with the Cells-to-CT technology. Figure 2. Real-time RT-PCR Using the TaqMan® MicroRNA Cells-to-CT™ Kit A dilution series of 10– HeLa cells was processed in triplicate with the TaqMan® MicroRNA Cells-to-CT™ Kit. miR was amplified from the. Catalog number: C. Cells-to-C T ™ Bulk Lysis Reagents are components of the TaqMan® Gene Expression Cells-to-C T ™ Kit, here made available as a separate purchase, in larger volumes than provided with the kit. The reagents included are: Lysis Solution, Stop Solution, and DNase.

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