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.