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FastLane Cell SYBR® Green Kit

For SYBR Green real-time one-step RT-PCR analysis from cultured cells without RNA purification

  • No RNA purification required, significantly saving time
  • Just 3 steps from cells to real-time RT-PCR
  • High-throughput analysis of 96- and 384-well plates
  • Unique gDNA Wipeout Buffer enables detection of RNA only
  • Immediate startup using optimized reagents and protocols

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).

Cat No./ID: 216213
FastLane Cell SYBR Green Kit (200)
€2,086.00
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FastLane Cell One-Step Buffer Set, 2x QuantiTect SYBR Green RT-PCR Master Mix, and QuantiTect RT Mix
The FastLane Cell SYBR® Green Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
0
Sensitive detection without RNA purification.
Two low-abundance transcripts, INPP5D (a phosphatase) and SRF (serum response factor), were quantified by real-time one-step RT-PCR. The FastLane Kit provided CT values comparable to those achieved with the RNeasy Mini Kit and QuantiTect SYBR® Green RT-PCR Kit.
1
Reliable detection of various transcripts.
Using the FastLane Kit, A549 cells in a 384-well plate were analyzed for the expression of Eg5, GAPDH, HMBS, and Rab5. Data show the mean CT value with standard deviation from triplicate wells. (Data kindly provided by Anne Grabner, Cenix BioScience GmbH, Germany.)
2
Highly specific quantification.
Real-time one-step RT-PCR using the FastLane Kit and a QuantiTect Primer Assay for INPP5D (a phosphatase) enabled specific transcript quantification in HeLa, HepG2, and Cos cells, as shown by the single peaks in melting curve analysis.
3
Highly reproducible analysis of a 96-well plate.
Real-time one-step RT-PCR using the FastLane Kit and a QuantiTect Primer Assay for INPP5D (a phosphatase) gave reproducible CT values from a 96-well plate of HeLa ACC cells.
Performance
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.
Principle

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.

Procedure

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.

Applications

The FastLane Cell SYBR Green Kit is ideal for experiments that require rapid, high-throughput gene expression analysis, such as:

  • Validation of siRNA-mediated gene knockdown
  • Screening of potential drugs
  • Detection of stimulated gene regulation
Features
Specifications
Applications SYBR Green-based, real-time, one-step RT-PCR
Enzyme activity Primer annealing
Mastermix Yes
Reaction type One-step, cell lysis, RNA stabilization, DNA digestion
Real-time or endpoint Real time
Sample/target type Cultured cells
Single or multiplex Single
With/without hotstart Without hotstart

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Kit Handbooks (3)
For real-time, one-step RT-PCR analysis of cultured cells without RNA purification
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RNA精製を行なわずに培養細胞から直接1ステップ・リアルタイムRT-PCR 解析
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