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qPCR Total Solution Seminar Series
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Dual-labeled fluorogenic probes are highly-sensitive, sequence-specific, fluorescent probes designed for real-time quantitative PCR1-3. They can be used with most real-time quantitative PCR instruments and multiplex analysis systems due to their simplicity of design and the extensive range of fluorophores available. |
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- Deprotected, desalted and purified by PAGE or RP-HPLC
- Available in lengths of 15 to 40 mers
- Delivered dried in individual, opaque tubes
- Shipped within 5 to 6 working days of receiving your order, pending successful QC validation
Guaranteed yields of dual-labeled fluorogenic probes
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Guaranteed yield (OD)
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Approximate yield (nmols*)
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1
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5
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5
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25
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10
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50
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50
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250
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100
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500
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*Estimate 1 OD = 5 nmols = 30 µg, for a 20 mer oligo
Please enquire for alternative quantities.
Labels for dual-labeled fluorogenic probes
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5'end reporter
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3'end quencher
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6-FAM™, HEX™, TET™, JOE™, TAMRA™, ROX™, Fluorescein, Cy™3, Cy5, Cy5.5, Texas Red™, Rhodamine, Rhodamine Red™, Rhodamine Green™, 6-Carboxyrhodamine 6G, Oregon Green™ 488, Oregon Green 500 or Oregon Green 514
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TAMRA, DABCYL, BHQ™-1 or BHQ-2
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Click here for a complete table of product specifications.
How do dual-labeled fluorogenic probes work?
A dual-labeled fluorogenic probe is a single-stranded oligonucleotide labeled with two different moieties: a reporter dye is located on the 5'end and an appropriate quencher is located on the 3'end. The quencher inhibits the natural fluorescence emission of the reporter dye by Förster-type energy transfer.
During the extension step of real-time quantitative PCR, the probe is excited by light from the PCR instrument (hγ1). The primer is extended and the Taq DNA polymerase 5'-3' exonuclease activity hydrolyzes the probe bound to the specific DNA template, releasing the reporter dye into solution where it is free to fluoresce and causing an increase in signal. The energy emitted by the reporter dye (hγ2) is detected by the real-time quantitative PCR system. The increase in measured fluorescent signal is directly proportional to the amount of accumulating target DNA.
Click here for reporters & quenchers available for bi-fluorescent probes.
References
1. Heid, C.A., et al. Real time quantitative PCR. Genome Research 6(10), 986-94, 2002.
2. Pusterla, N., et al. Quantitative real-time PCR for detection of members of the Ehrlichia phagocytophila genogroup in host animals and Ixodes ricinus ticks. J. Clin. Microbiol. 37(5), 1329-31, 1999.
3. Saito T., et al. Quantitative DNA analysis of low-level hepatitis B viremia in two patients with serologically negative chronic hepatitis B. J. Med. Virol. 58(4), 325-31, 1999.
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