Description |
The 2x One-tube RT-PCR Hot Start kit is designed for combining two reactions of reverse transcription and PCR. It provides more simple and effective operation in RT-PCR. The reverse transcription step is working by M-MLV RT (H-). The PCR reaction is working by Hot start Taq DNA Polymerase. An especially reaction buffer is provided both for Reverse Transcriptase and Hot Start Taq DNA Polymerase.
Storage condition |
long time at -20°C
Mix component |
- M-MLV RTase
- Hot start Taq
- RT-PCR reaction buffer
- dNTP
- stabilizer
Sensitivity |
Targets can generally be detected from < 1 pg to 50 ng polyA RNA(mRNA) or 10 pg to 1 µg total RNA. Even lower amounts of RNA may be successfully amplified by using highly expressed transcripts.
Cat No |
Pack size |
| TAMB17Z-50 | 50 rx |
| TAMB17Z-100 | 100 rx |
| TAMB17Z-500 | 500 rx |
PCR cycles program |
| Step | Temperature | Time | Cycle |
| RT reaction | 37-50ºC | 30-120 mins | 1 |
| Initial denaturation | 94ºC | 10 mins | 1 |
| Denaturation | 94ºC | 0.2-1 sec | 30-45 |
| Annealing | 50-68ºC | 0.2-1 sec | |
| Extension | 72ºC | 1min/1kb | |
| Final extension | 72ºC | 1-10 mins | 1 |
IMPORTANT: Annealing temperature should be 2-6ºC lower than the primer melting temperature.
Analyzing the RT-PCR Products |
Analyze the RT-PCR products by 1.0% (w/v) agarose gel electrophoresis. The products will be visible by UV transillumination of the ethidium bromide-stained agarose gel. Store the reaction products at –20°C until needed.
