FastLane Cell Probe Kit (200)

From QIAGEN, this kit enables sensitive probe-based detection in real-time one-step RT-PCR directly from cultured cells, eliminating the need for RNA purification. The workflow combines cell lysis with integrated RNA stabilization and genomic DNA elimination, after which lysates can be used directly in the RT-PCR reaction. Real-time quantification is reported as more sensitive and linear compared with similar kits, with detection reported down to a single cell and high reproducibility across multiwell plate formats. Genomic DNA does not interfere with quantification.

All probe chemistries are compatible, including hydrolysis probes (such as TaqMan or other dual-labeled probes), LightCycler hybridization (FRET) probes, and Molecular Beacons. The ready-to-use RT-PCR master mix is described as containing a novel RT-PCR buffer and a hot-start DNA polymerase, removing the need to optimize probe concentration or other PCR parameters. A three-step lysate preparation procedure takes approximately 12 minutes (cell wash, cell lysis with RNA stabilization and DNA elimination, and lysate pretreatment), after which lysates can be used immediately or stored at -80°C for up to 32 months and subjected to up to three freeze-thaw cycles.

The kit contains the FastLane Cell One-Step Buffer Set, 2x QuantiTect Probe RT-PCR Master Mix, and QuantiTect RT Mix. It has been successfully tested on suspension cells, primary cells, and cell lines including HeLa, Jurkat, HUVEC, HepG2, UASMC (umbilical artery smooth muscle cells), MCF7, and NHBE (normal human bronchial epithelial cells). The product is intended for molecular biology research applications such as rapid high-throughput gene expression analysis.