FastLane Cell Probe Kit
For probe-based real-time one-step RT-PCR analysis from cultured cells without RNA purification
The FastLane Cell Probe Kit accelerates and streamlines real-time RT-PCR and is ideal for experiments requiring rapid high-throughput gene expression analysis.The kit provides a set of buffers for preparing FastLane lysates from cultured cells. In addition to lysing cells, the buffers also stabilize cellular RNA and eliminate genomic DNA. FastLane lysates are used directly in probe-based real-time one-step RT-PCR. A variety of other detection formats are also available, including detection with SYBR Green or multiplex detection with sequence-specific probes (with or without ROX passive reference dye).
The FastLane Cell Probe Kit delivers sensitive probe-based detection in real-time one-step RT-PCR without the need for RNA purification (see figure "Sensitive probe detection without RNA purification"). Real-time quantification is more sensitive and linear compared with similar kits (see figures "Greater sensitivity than other kits" and "Linear detection down to a single cell"), and is highly reproducible (see figure "Highly reproducible analysis of a 384-well plate"). In addition, genomic DNA does not interfere with quantification.
The FastLane Cell Probe Kit uses FastLane technology to prepare cell lysates for real-time one-step RT-PCR. All types of probe chemistry are compatible with the FastLane Cell Probe Kit, including hydrolysis probes (e.g., TaqMan® or other dual-labeled probes), LightCycler hybridization (FRET) probes, and Molecular Beacons. A ready-to-use RT-PCR master mix containing a novel RT-PCR buffer and a hot-start DNA polymerase eliminates the need to optimize probe concentration and other PCR parameters.
With the FastLane Cell Probe Kit, lysates are obtained in just 12 minutes in a 3-step procedure: cell wash; cell lysis with integrated RNA stabilization and genomic DNA elimination; and lysate pretreatment. After pretreatment, lysates are ready to use in real-time one-step RT-PCR.
The kit contains sufficient reagents for cell lysis in two 96-well cell culture plates and subsequent real-time RT-PCR (50 µl PCR volume). The kit can also be used to analyze a single 384-well cell culture plate if the PCR volume is reduced to 20 µl; if a 50 µl PCR volume is desired, an additional QuantiTect Probe RT-PCR Kit (200) needs to be purchased.
The FastLane Cell Probe Kit is ideal for experiments which require rapid, high-throughput gene expression analysis, such as:
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