Quantitative real-time PCR (qPCR) probes

The Real-time PCR (qPCR) or 5'-Nuclease assay is a sensitive assay for online monitoring of amplification kinetics. Real-time PCR Probes consist of an oligonucleotide complementary to the amplified probe. The probe is labeled at the 5'-end with the fluorescence donor (usually Fluorescein, HEX or TET) and a few bases downstream or on the 3'-end with a quencher, usually TAMRA. When no complementary sequence is available, the fluorescence of the donor is quenched. During PCR the labeled oligonucleotide hybridizes to the target sequence and the 5'-dye is removed by 5'3'-exonuclease activity of Taq. The fluorescence of the donor is no longer quenched and can be measured. The fluorescence intensity is proportional to the amount of PCR-product formed during the early exponential phase of PCR (threshold value).

Real-time PCR principle
Real-time PCR principle


In the probe design it is important that the melting point of the probe is higher than that of the upstream primer. The 3'-end of the probe must be protected against chain elongation during PCR. For blocking the 3'-end phosphate, cordycepin, 2',3'-dideoxynucleosides, inverseT or the quencher dye itself can be used. Labeling the 3'-end with TAMRA or another quencher can be achieved by using a 3'-amino-linker. Labeling internal positions of the sequence is possible by incorporating a modified thymidine residue which contains an amino-link at the 5-position of the base. This attachment does not interfere with hybridization.

We offer

  • great variety of oligonucleotides with different labels and their combinations (see our list of dyes)
  • 5´- , 3´- or internal labels
  • Click chemistry for flexible dual labeling
  • high purity probes and thorough quality control

We offer a large selection of dye combinations suitable for Real-time PCR probes. Please make a request if your combination of interest is not listed below! 

Product

Amount

Cat. no.

Price/coupling

Real-time PCR probe

5' FAM 3' TAMRA

5 nmol

5-0901-002

142.00 EUR

25 nmol

5-0901-003

184.00 EUR

100 nmol

5-0901-004

347.00 EUR

Real-time PCR probe

5' HEX 3' TAMRA

5 nmol

5-0901-012

194.00 EUR

25 nmol

5-0901-013

268.00 EUR

100 nmol

5-0901-014

446.00 EUR

Real-time PCR probe

5' TET 3' TAMRA

5 nmol

5-0901-022

194.00 EUR

25 nmol

5-0901-023

268.00 EUR

100 nmol

5-0901-024

373.00 EUR

To order your tailor-made oligo above get in contact with our individual personal assistance provided by nucleic acid chemists and molecular biologists. Your personal contact person from IBA Lifesciences is ready to help you by email or phone (+49 551 50672-222) and will support you in designing your correct oligo sequence and advise you in possible promising labeling and modification combinations. Furthermore our assistance alerts you to create incorrect not functional oligonucleotides before starting the synthesis.