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Description:
The One-step Q-PCR Kit utilizes a proprietary hot-start
PCR technology. It enables sensitive detection of RNA and fast
thermocycling.
Application:
This reaction mix DOES NOT contains internal passive reference
dye (ROX). It is manufactured for the following real-time PCR
machines: ABI 5700, 7000, 7300, 7900HT. Due to a different
optical system, it is recommended ABI 7500 users use product
# G124.
Recommended storage conditions:
This product should be stored at –20ºC. To avoid repeated
freeze-thaw, open vials should be kept at 4ºC.
Primer and probe design:
Appropriate software, such as ABI Primer Express™, should be
used.
1. Tm: 58~60°C with 17~25 nucleotides in length
2. To design a small amplicon, <150bp
3. To avoid secondary structures
4. To avoid more than 3 consecutive Gs
5. To avoid complementary 3’ ends
Recommended reaction conditions:
55°C, 5min -> 95°C, 5min. -> (95°C, 5sec. -> 60°C, 30sec.) for
50 cycles.
Tips to get good performance with One-Step RT PCR
- Special caution should be taken to avoid any RNase
contamination
- To design probe on sense strand
- Primer concentration: 100nM to 300nM
- To shorten time between setting up reaction and loading plate
onto PCR machine
Reference:
Holland, P. M., Abramson, R. D., Watson, R., and Gelfand, D. H.
1991. Proceedings of the National Academy of Sciences USA
88:7276–7280
This product is distributed
for laboratory research only.
Caution: Not for diagnostic use . |
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