DNA Safe Stains
2X One-tube RT-PCR mix
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
1. M-MLV RTase
2. Hot start Taq
3. RT-PCR reaction buffer
4. dNTP
5. 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 | 37-50°C | 30-120 mins | 1 |
| Initial denaturation | 94°C | 10 mins | 1 |
| Denaturation | 94°C | 0.2-1 sec | 30-45 |
| Annealing | 50-68°C | 0.2-1 sec | |
| Extension | 72°C | 1min/1kb | |
| Final extension | 72°C | 1-10 mins | 1 |
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.
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