FastLane Cell SYBR® Green Kit
For SYBR Green real-time one-step RT-PCR analysis from cultured cells without RNA purification
The FastLane Cell SYBR Green 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 SYBR Green-based real-time one-step RT-PCR. A variety of other detection formats are also available, including singleplex detection with sequence-specific probes or multiplex detection with sequence-specific probes (with or without ROX passive reference dye).
The FastLane Cell SYBR Green Kit uses a unique RT-PCR buffer to prevent the formation of nonspecific products and primer–dimers, allowing highly specific quantification in real-time one-step RT-PCR (see figure "Highly specific quantification"). Sensitive quantification is also achieved despite the absence of an RNA purification step (see figure "Sensitive detection without RNA purification"). Results in high-throughput experiments are very reproducible (see figures "Highly reproducible analysis of a 96-well plate" and "Reliable detection of various transcripts") and not affected by genomic DNA contamination.
The FastLane Cell SYBR Green Kit uses FastLane technology to prepare cell lysates for real-time one-step RT-PCR. The kits accelerate and streamline real-time PCR analysis of cultured cells by eliminating the need for RNA purification and deliver reproducible results in high-throughput analysis, enabling easy processing of 96 or 384-well plates.
Specific transcript detection is guaranteed when the kit is used in combination with QuantiTect Primer Assays. These are predesigned, genomewide primer sets, available at GeneGlobe.
With the FastLane Cell SYBR Green 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 SYBR Green RT-PCR Kit (200) needs to be purchased.
The FastLane Cell SYBR Green Kit is ideal for experiments that require rapid, high-throughput gene expression analysis, such as:
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