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FastLane Cell cDNA Kit (50)
Cat. No. / ID: 215011
Buffer FCW, Buffer FCP, and components for 50 x 20 µl reverse-transcription reactions (gDNA Wipeout Buffer, Quantiscript Reverse Transcriptase, Quantiscript RT Buffer, RT Primer Mix, and RNase-Free Water)
The FastLane Cell cDNA Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
Features
- From cells to cDNA in only 4 steps and under 45 minutes
- Sensitive detection of even low-abundance transcripts
- Easy parallel processing of several samples
- Integrated gDNA removal allows detection of RNA only
- No need to design RNA-specific primers or probes
Product Details
The FastLane Cell cDNA Kit provides a fast and simple procedure for preparing first-strand cDNA directly from cultured cells without RNA purification. The kit is supplied with wash and lysis buffers for preparing lysates and stabilizing RNA, gDNA Wipeout Buffer for eliminating genomic DNA contamination, and all reaction components for fast and efficient cDNA synthesis. The FastLane Cell cDNA Kit is ideal for experiments requiring rapid, high-throughput gene expression analysis.
Performance
cDNA synthesized by the FastLane Cell cDNA Kit enables highly sensitive and reproducible results in real-time two-step RT-PCR (see figures " Superior sensitivity in cancer research" and " Highly reproducible cDNA preparation"). The high quality of the cDNA also allows detection over a wide linear range from >105 cells down to one cell.
See figures
Principle
The FastLane Cell cDNA Kit uses FastLane technology to prepare cDNA directly from cultured cells in just 45 minutes. To ensure only RNA is detected in real-time RT-PCR, novel gDNA Wipeout Buffer is used to eliminate genomic DNA (see figure " Effective elimination of genomic DNA contamination"). Time and effort are saved as there is no need to design RNA-specific primers or probes.
When the FastLane Cell cDNA Kit is used together with optimized QuantiTect, QuantiFast, or Rotor-Gene Kits for real-time RT-PCR, several cell samples can be easily processed and analyzed within a few hours. This allows a snapshot of several transcripts in experiments such as validation of siRNA-mediated gene knockdown (see figure " Successful analysis of CDC2 knockdown").
The FastLane Cell cDNA Kit is part of the comprehensive range of FastLane Kits, which speed up and streamline the gene expression analysis workflow.
See figures
Procedure
With the FastLane Cell cDNA Kit, cDNA is obtained from cultured cells in just 45 minutes in a 4-step procedure: cell wash; cell lysis with integrated RNA stabilization; genomic DNA elimination; and cDNA synthesis. The synthesized cDNA is ready to use in real-time two-step RT-PCR.
The kit contains sufficient reagents for preparing cDNA from two 24-well cell-culture plates or one 48-well cell-culture plate. If working with a 96-well cell-culture plate, an additional QuantiTect Reverse Transcription Kit (50) needs to be purchased.
cDNA prepared with the FastLane Cell cDNA Kit can be analyzed by real-time two-step RT-PCR using one of the following QuantiTect Kits:
- SYBR® Green detection: QuantiTect SYBR® Green PCR Kit
- Probe detection: QuantiTect Probe PCR Kit
- Multiplex probe detection: QuantiTect Multiplex PCR Kits
Alternatively, fast real-time two-step RT-PCR can be carried out when the FastLane Cell cDNA Kit is used in combination with a QuantiFast Kit:
- SYBR® Green detection: QuantiFast SYBR® Green PCR Kit
- Probe detection: QuantiFast Probe PCR Kits
- Multiplex probe detection: QuantiFast Multiplex PCR Kits
Ultrafast real-time two-step RT-PCR can be achieved when cDNA prepared with the FastLane Cell cDNA Kit is analyzed on the Rotor-Gene Q cycler using one of the following dedicated Rotor-Gene Kits:
- SYBR® Green detection: Rotor-Gene SYBR® Green PCR Kit
- Probe detection: Rotor-Gene Probe PCR Kit
- Multiplex probe detection: Rotor-Gene Multiplex PCR Kit
QuantiTect and QuantiFast Kits are compatible with various real-time cyclers, including instruments from Applied Biosystems.
Applications
The FastLane Cell cDNA Kit is suitable for experiments which require a snapshot of individual transcript levels, such as:
- Validation of siRNA-mediated gene knockdown
- Evaluation of drug effects
- Detection of gene regulation
Supporting data and figures
Superior sensitivity in cancer research.
In real-time two-step RT-PCR analysis of various genes important to cancer research in HeLa cells, lower CT values were achieved when cDNA was prepared with the FastLane Kit instead of a similar kit from Supplier AIV.
Specifications
| Features | Specifications |
| Applications | Real-time, two-step RT-PCR, gene expression analysis directly from cells |
| With/without hotstart | Without hotstart |
| Enzyme activity | Reverse transcription |
| Reaction type | cDNA production, DNA digestion |
| Mastermix | No |
| Real-time or endpoint | Real-time, two-step RT-PCR |
| Sample/target type | Cultured cells |
| Single or multiplex | Single |
Resources
Kit Handbooks (1)
Safety Data Sheets (1)
Brochures & Guides (1)
FAQ
Is there a stopping point in the protocol for Suspended Cells using the FastLane Cell cDNA Kit?
Can the FastLane Cell cDNA Kit also be used for qualitative end-point PCR?
Is it possible to use frozen cell pellets with the FastLane Cell cDNA Kit?
Can I use my gene-specific primers with the FastLane Cell cDNA Kit and the QuantiTect Reverse Transcription Kit?
How do the FastLane Cell cDNA Kit and QuantiTect Reverse Transcription Kit eliminate genomic DNA contamination?
How many cells can be used with the FastLane Cell cDNA Kit?
What cell lines have been tested with the Fastlane Cell cDNA Kit?
Will the FastLane Cell cDNA Kit work with suspension cells?
Can I use specific primers with the FastLane Kits?
Can the FastLane RT reaction be incubated longer than 30 min at 42 deg C?
Can the FastLane Cell lysates (containing stabilized RNA, step 8. in Fastlane protocol) be stored?
What is the cell number range one can use with the FastLane Cell cDNA Kit for real-time RT-PCR?
Can the FastLane Cell cDNA Kit be used with tissue?