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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:
Contains all components for one-step Q-PCR, includes passive
reference dye (ROX) enable signal normalization in all 5’
nuclease assays, quantify RNA targets, enhanced performance,
minimizing setup time. It is manufactured for the ABI 7500 PCR
machine.
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 Q- 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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