Detection
Fluorogenic probes allow real-time quantitative detection of nucleic acid sequence amplification(1-5).
Bilabeling
These probes are bilabeled fluorescent oligonucleotides with a 5'-end reporter dye (6-Fam, Hex or Tet) and a 3'-end quencher dye (Tamra).
At close proximity, the quencher dye suppresses the characteristic fluorescence emission of the reporter dye.
Hybridization
Fluorogenic probes hybridize with the target DNA sequence in a region located between the forward and reverse PCR* primer hybridization sites.
During PCR*, the 5'-3' Taq polymerase exonuclease activity releases the reporter dye, causing fluorescence.
Applications
The Fluorogenic Probes are ideally suited for large-scale and high-throughput target sequence detection applications, such as(2-5):
- quantitative gene analysis
- large-scale association mapping
- automated allelic discrimination
- large-scale mutation screening
- high-throughput haplotyping
- automated pathogen detection
Sequence
Sequence Probe should not contain any secondary structure (hairpin, self-hybridization)
and should not hybridize or overlap with the primers.
Quality Entirely deprotected and desalted. Purified by reverse phase HPLC.
Length optimal size: 20 to 40 bases.
At least 5℃ higher than the Tm of the primers to increase the stability
of the probe on the target DNA.
Fluorophores
Fluorophore 5'-end reporter (6-Fam, Hex, or Tet).
Fluorophore 3'-end quencher (Tamra).
Quantity 10 OD, 5 OD, 1 OD. For larger quantities, please contact us.
Quality Controls
Length and quality by PAGE analysis.
Purified by reverse phase HPLC.
Labels' absorbance profiles.
Form
Form Delivered in a highly concentrated, ready-to-use distilled water solution.
Guarantees
Conformity of the sequence.
Minimum final quantity.
Quality according to above specifications.
Documentation
1 - Oligonucleotide Technical Data Sheet
- Concentration (μM and μg/μl), quantity (OD and nmoles), Tm, MW, size and molecular extinction coefficient.
2 - Analysis Certificate
- Reverse phase HPLC profile at 260 nm
- Labels' absorbance measurements.
Turn-around time
5 working days.
* PCR is patented by Hoffman La Roche Ltd.
* For research use only.
References
1. Livak, K.J., et al. (1995). Oligonucleotides with fluorescent dyes at opposite ends provide
a quenched probe system useful for detecting
PCR product and nucleic acid hybridization.
PCR Methods Appl. 4(6): 357-362.
2. Heid, C.A., et al. (1996). Real time quantitative PCR. Genome Res. 6(10): 986-994.
3. Gelmini, S., et al. (1997). Quantitative polymerase chain reaction-based homogeneous assay
with fluorogenic probes to measure c-erbB-2 oncogene
amplification. Clin Chem. 43(5): 752-758.
4. Pusterla, N., et al. (1999). 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.
5. Saito, T., et al. (1999). 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.
